CN106093430A - Can be used for mark detecting diabetes and application thereof - Google Patents
Can be used for mark detecting diabetes and application thereof Download PDFInfo
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- CN106093430A CN106093430A CN201610393254.3A CN201610393254A CN106093430A CN 106093430 A CN106093430 A CN 106093430A CN 201610393254 A CN201610393254 A CN 201610393254A CN 106093430 A CN106093430 A CN 106093430A
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- G—PHYSICS
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- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
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- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6803—General methods of protein analysis not limited to specific proteins or families of proteins
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- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6803—General methods of protein analysis not limited to specific proteins or families of proteins
- G01N33/6848—Methods of protein analysis involving mass spectrometry
- G01N33/6851—Methods of protein analysis involving laser desorption ionisation mass spectrometry
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- G01N2800/00—Detection or diagnosis of diseases
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Abstract
The present invention is provided to determine whether experimenter has or by Developing Grape impaired glucose tolerance maybe by the mark of development type 2 diabetes mellitus, comprising: one or more in glucose, β mannose monoglyceride, D talose, diagnostic sensitivity is up to 96.3%, and specificity is 0%.
Description
Technical field
The present invention relates to a kind of diabetes mark, especially, be specifically related to G01N33/68 further.
Background technology
The pathophysiological basis occurring glucose metabolism to change is multifactorial, such as life style and inherited genetic factors.
Especially, in heredity in the people of susceptible type 2 diabetes mellitus, obesity is considered as the main cause of type 2 diabetes mellitus, and in the past
Between 50 years, the ratio of type 2 diabetes mellitus dramatically increases abreast with obesity.Compare about 0.3 hundred million people of 1985, by 2010 be
Only, about 2.85 hundred million people suffer from this disease.
Develop into type 2 diabetes mellitus from NGT (Healthy People) and include impaired fasting plasma glucose (IFG) and Fructus Vitis viniferae
The interstage of impaired glucose tolerance (IGT), referred to as prediabetes.
Although such as age, Body Mass Index (BMI), the generation to prediabetes and type 2 diabetes mellitus is relevant, but these because of
Element is not enough to Accurate Prediction and develops into the risk of impaired glucose tolerance from Healthy People and/or develop into from impaired glucose tolerance
The risk of type 2 diabetes mellitus, because the development of diabetes and progress are the most asymptomatic.Although additionally, being used for determining experimenter
The method whether with impaired glucose tolerance and/or type 2 diabetes mellitus is known, but such method require overnight fast and
Repeatedly draw blood within a few hours, and be often accompanied by side effect, such as Nausea and vomiting, abdominal bloating and/or headache.
Because to there is the experimenter of impaired glucose tolerance and/or type 2 diabetes mellitus and/or there is Developing Grape carbohydrate tolerance
The experimenter of the risk of impaired and/or type 2 diabetes mellitus and/or the early stage of those experimenters that specific therapy responds will be identified
The short-term relevant to glucose disorders and long-term complications will be reduced, so this area is it needs to be determined which experimenter has or incites somebody to action
Developing Grape impaired glucose tolerance and/or type 2 diabetes mellitus and/or respond a certain therapy reliable and accurately method, to permit
Permitted early intervention.
Summary of the invention
The present invention is based at least partially on relevant to the development of impaired glucose tolerance and/or type 2 diabetes mellitus and has
The discovery of relevant mark is reacted in treatment by the experimenter of impaired glucose tolerance and/or type 2 diabetes mellitus.Therefore, this
Bright by measuring and identify that specific mark or specific mark combine, it is provided that to be used for predicting whether experimenter has or incite somebody to action
Sensitive and the easy way of Developing Grape impaired glucose tolerance and test kit, be used for predicting whether experimenter has maybe by sugared for development
The method of urine disease and test kit, and for identifying the chemical combination of the progress that can slow down impaired glucose tolerance and/or type 2 diabetes mellitus
The method of thing, monitors a certain therapy effect in the progress of the impaired glucose tolerance and/or type 2 diabetes mellitus that lower experimenter
Method, and and/or the method for type 2 diabetes mellitus progress impaired for suppression glucose is resistance in cell or experimenter amount.
Therefore, in one aspect, the present invention is provided to determine whether experimenter has or be subject to by Developing Grape carbohydrate tolerance
The mark damaged: glucose, β-mannose monoglyceride, D-talose.
Described mark may further comprise: cystine, sorbose, agedoite, lysine, isoleucine, threonine,
One in fumaric acid, hydroxyurea, xylose, tyrosine, leucine, aspartic acid, glyceryl monostearate, linoleic acid, glycerol
Or it is multiple.
On the one hand, the present invention is provided to determine experimenter whether have or by Developing Grape impaired glucose tolerance and/or
By development type 2 diabetes mellitus and/or by the method for development type 2 diabetes mellitus related complication, described method includes:
The sample of experimenter measures above-mentioned in the level of one or more marks；
By the level of one or more marks in Samples subjects and one or more marks in control sample
Level compare, the one wherein compared with the level of one or more marks in control sample, in Samples subjects
Or the difference of the level of multiple markers indicates described experimenter to have or by Developing Grape impaired glucose tolerance.
On the other hand, the present invention is provided to determine the experimenter with impaired glucose tolerance and/or type 2 diabetes mellitus
The method that whether therapy will be responded, described method includes:
The level of one or more marks above-mentioned is measured in the sample of experimenter；
By the level of one or more marks in Samples subjects and one or more marks in control sample
Level compare, the one wherein compared with the level of one or more marks in control sample, in Samples subjects
Or the difference of the level of multiple markers indicates described experimenter will respond described therapy.
On the other hand, the present invention is provided to suppress the impaired glucose tolerance of experimenter and/or sending out of type 2 diabetes mellitus
The method of exhibition.Described method includes the reagent giving described experimenter's effective dose, any one of the described reagent regulation present invention
Or the expression of multiple markers and/or activity, thus suppress the impaired glucose tolerance of experimenter and/or sending out of type 2 diabetes mellitus
Exhibition.
In one embodiment, in Samples subjects horizontally through mass spectrometric determination.In one embodiment, institute
Stating mass spectrography is laser desorption/flight time (MALDI/TOF) mass spectrography of Matrix-assisted, liquid chromatograph quadruple ion capture electricity
Injection (LCQ-MS) or Protein-based tumor biomarker/flight time (SELDI/TOF) mass spectrography.
In another embodiment, measuring horizontally through immunoassay in Samples subjects.
The sample of experimenter can be fluid sample or tissue sample.
In one embodiment, the level of described mark is expression and/or the activity of mark.
In one embodiment, described experimenter has the risk of development type 2 diabetes mellitus.
In one aspect, the present invention is provided to determine whether experimenter has or by Developing Grape impaired glucose tolerance or general
Development type 2 diabetes mellitus is maybe by the test kit of development type 2 diabetes mellitus complication, and described test kit includes in Samples subjects
The reagent of the level of one or more marks in claim 1 and determine whether experimenter has or by resistance to for Developing Grape sugar
Measure impaired maybe by development type 2 diabetes mellitus maybe will development type 2 diabetes mellitus complication test kit operation instructions.
On the other hand, the present invention is provided to determine the experimenter with impaired glucose tolerance and/or type 2 diabetes mellitus
The test kit that whether treatment will be responded, described test kit include for measure in Samples subjects above-mentioned in one or
The reagent of the level of multiple markers and determine the test kit operation instructions whether experimenter will respond to described treatment.
Monitor in there is the experimenter of impaired glucose tolerance and/or type 2 diabetes mellitus it yet still another aspect, the present invention provides
The test kit of effect for the treatment of.Described test kit includes the level for measuring one or more marks in Samples subjects
Reagent, and monitor the test kit operation instructions of effect of described treatment.
In one embodiment, described test kit farther includes the reagent for obtaining sample from experimenter.
In one embodiment, described test kit farther includes control sample.
In one aspect, the method that the present invention is provided to identify type 2 diabetes mellitus mark, described method includes:
Under steady-state conditions, identify the blood of two or more from two or more species, thus produce
The temporary table of mark；
Mark in temporary table is identified under the conditions of dysfunction；
Identify mark in temporary table under normal operation；
The level of the mark in the sample of determination test sample and control sample, wherein with the level phase in test specimen
Ratio, in control sample, mark described in the differential identification of the level of mark is type 2 diabetes mellitus biomarker；
Wherein test specimen is from having the experimenter of impaired glucose tolerance or from being newly diagnosed as being subject to of type 2 diabetes mellitus
Examination person or from the experimenter having been determined as type 2 diabetes mellitus；
Wherein control sample is from having the experimenter of NGT or from having impaired glucose tolerance
Experimenter or from the experimenter being newly diagnosed as type 2 diabetes mellitus.
Accompanying drawing is sketched
Fig. 1: embodiment 1, schemes with cystine for ROC during mark；
Fig. 2: embodiment 2, schemes with glucose for ROC during mark；
Fig. 3: embodiment 10, schemes with isoleucine for ROC during mark；
Fig. 4: embodiment 20, schemes with cystine and glycerol for ROC during mark；
Fig. 5: embodiment 30, schemes with glucose and D-talose for ROC during mark；
Fig. 6: embodiment 40, schemes with cystine, glucose and glycerol for ROC during mark；
Fig. 7: embodiment 50, schemes with aspartic acid, glucose and glycerol for ROC during mark；
Fig. 8: embodiment 70, with cystine, aspartic acid, β-mannose monoglyceride and glycerol for ROC during mark
Figure；
Fig. 9: embodiment 55, schemes with glucose, β-mannose monoglyceride and D-talose for ROC during mark；
Figure 10: principal component analysis figure, wherein circle represents healthy group, and rectangle represents diabetic groups.
Detailed description of the invention
The present invention is based at least partially on and impaired glucose tolerance and/or the development of type 2 diabetes mellitus, type 2 diabetes mellitus
It is in progress and there is the experimenter of impaired glucose tolerance and/or type 2 diabetes mellitus to treating the discovery responding relevant mark.
Therefore, the present invention is by measuring and identify that specific mark or specific mark combine, it is provided that be used for predicting
Experimenter whether have or by method and the test kit of Developing Grape impaired glucose tolerance, be used for predicting whether experimenter has or will
The development method of diabetes and test kit and for identifying the progress of can slow down impaired glucose tolerance and/or type 2 diabetes mellitus
The method of compound, monitoring therapy merit in reducing the progress of the impaired glucose tolerance of experimenter and/or type 2 diabetes mellitus
Effect method and for suppression in cell or experimenter the method for the progress of impaired glucose tolerance and/or type 2 diabetes mellitus.
Various aspects of the invention are described in greater detail in following sections:
Definition
As used herein, each of following term has the implication relevant in this part to it.
Article " one " and " a kind of " are used for referring to one of this article or language more than one (i.e. at least one) herein
Method object.Such as, " key element " means a key element or more than a key element.
" mark " or " biomarker " is compared with another kind of phenotypic status (such as not suffering from disease), is taking from one
The organic biomolecules that in the sample of the experimenter planting phenotypic status (such as suffering from disease), difference exists.If in different groups
Average or the Median levels of biomarker, such as expression are calculated as statistically significant, then biomarker is not
In same phenotypic status, difference exists.The common inspection of significance,statistical is particularly including t-inspection, ANOVA, Kruskal-
Wallis, Wilcoxon, Mann-Whitney and odds ratio.Biomarker alone or in combination provides experimenter to belong to a kind of
The tolerance of the relative risk of phenotypic status or another kind of phenotypic status.Therefore, they are used as mark, such as, (pre-for disease
Afterwards and diagnosis), the therapeutic efficiency of medicine (treatment diagnostics) and the mark of drug toxicity.
In some embodiments, the accuracy for the mark of the compositions and methods of the invention can pass through receptor
Performance curve (" ROC curve ") characterizes.ROC is the True Positive Rate phase of the cutpoint that the difference for diagnosis marker is possible
Curve to false positive rate.Relation between ROC curve display susceptiveness and specificity.I other words, the increase of susceptiveness will companion
With specific reduction.Curve is the closer to the left axle in ROC space, then apical margin, and mark is the most accurate.On the contrary, curve more leans on
45 degree of diagonal of nearly ROC figure, mark is the most inaccurate.Area under ROC is the tolerance of mark accuracy.The standard of mark
Really property depends on that group to be tested is the most suitably divided into the group with described disease and does not have described disease by mark
Group.Area under curve (referred to as " AUC ") is the 1 perfect mark of expression, and area is the mark that 0.5 expression serviceability is poor
Thing.Therefore, in some embodiments, the biomarker of the present invention and the AUC of method be greater than about 0.50, greater than about 0.60 or
Greater than about 0.70.
" type 2 diabetes mellitus " is also known as " diabetes " in this article, is characterized as peripheral insulin resistance and pancreatic beta cell not
The sufficiently combination of insulin secretion.If the fasting glucose of experimenter (FPG) level is about 126mg/dL (about 7.0mmol/L)
Or it is higher；It is about 200mg/dL (about in 75-g oral glucose tolerance test (OGTT) period 2-hours blood glucose (PG) level
11.1mmol/L) or higher；In having the experimenter of symptom of hyperglycemia or hyperglycemia crisis, random blood sugar is about
200mg/dL (about 11.1mmol/L) or higher；And/or glycated hemoglobin (HbA1c) level is about 6.5% or higher, then " it is subject to
Examination person has diabetes ".
There is the experimenter of " NGT " or " NGT " in 75-g oral glucose tolerance test (OGTT) period
2-hours blood glucose (PG) level is less than about 140mg/dL (less than about 7.8mmol/L)；Fasting glucose (FPG) level is for less than about
110mg/dL (less than about 6.1mmol/L)；And/or glycated hemoglobin (HbA1c) level is less than about 6%.
" having the experimenter of the risk of development diabetes " is such experimenter, and its lasting blood pressure is about 135/80mmHg
Or it is higher；It is overweight that (such as Body Mass Index (BMI) is greater than about 30kg/m2)；Relevant to diabetes first degree；HDL level is about
35mg/dL or higher and/or triglyceride levels are less than about 250mg/dL；Age is 45 years old or bigger；It is women；There is gestation
Diabetic history；There is polycystic ovary syndrome；There is the patient's condition relevant to metabolism syndrome；It is Spaniard；It is that Africa nationality is beautiful
Compatriots；And/or be Native Americans.Additionally, many Drug therapys and Other diseases may make experimenter have development diabetes
Risk.Such as, glucocorticoids, thiazide, beta-blocker, atypical antipsychotic and statins may make
Experimenter has the risk of development diabetes.Had suffered from before acromegaly, hyperthyroidism, pheochromocytoma and certain
The experimenter that a little cancer such as glucagonomas of pancreas and testosterone lack also has the risk of development type 2 diabetes mellitus.
Experimenter, such as, have the experimenter of development diabetes risk, it may be possible to " prediabetic ".If experimenter
There is impaired glucose tolerance, then it is assumed that described experimenter is " prediabetic "." impaired glucose tolerance " is hyperglycemia
A kind of state of disease, it is relevant with insulin resistant and cardiovascular pathology risk increase.After when 2 is little, experimenter has middle liter
High glucose level, but less than the level limited for type 2 diabetes mellitus, then experimenter has impaired glucose tolerance.Empty
Abdomen blood glucose is probably normal or slightly elevated.
In 75-g oral glucose tolerance test (OGTT) period, there is 2-hour of experimenter of impaired glucose tolerance
Blood glucose (PG) level is about 140mg/dL (about 7.8mmol/L) or higher (such as about between 7.8 and 11mmol/L)；Fasting blood
Sugar (FPG) level is less than about 126mg/dL (less than about 7mmol/L) (such as between about 95 and about 125mg/dL)；Blood red egg
White A1c (HbA1c) level is about 6% or higher (such as about between 6.0 and 6.4)；And/or BMI is about 24kg/m2Or more
Greatly.
Experimenter, such as, have the experimenter of development diabetes risk, be likely to be of " glucemia is impaired on an empty stomach ".At 75-g mouth
Take glucose tolerance test (OGTT) period, there is 2-hours blood glucose (PG) level of the experimenter that glucemia is impaired on an empty stomach for being less than
About 140mg/dL (less than about 7.8mmol/L)；Fasting glucose (FPG) level is less than about 126mg/dL (less than about 7mmol/L)
(such as between about 110 and about 125mg/dL)；And/or glycated hemoglobin (HbA1c) level is about 6% or higher and (such as exists
About between 6.0 and 6.4).
Term " diabetes develop " refers in experimenter, and it is impaired that NGT develops into glucemia on an empty stomach；Just
Often glucose tolerance develops into impaired glucose tolerance；NGT develops into type 2 diabetes mellitus；Impaired of glucemia on an empty stomach
Exhibition is impaired glucose tolerance；Glucemia is impaired on an empty stomach develops into type 2 diabetes mellitus；And/or impaired glucose tolerance develops into 2 type sugar
Urine disease.
" level of mark " or " level of biomarker " refers to mark present in the sample to be tested
Amount.The level of mark can be abswolute level or amount (such as μ g/ml) or relative level or measure (such as relative signal intensity).
" higher level " or " level increase " of mark refers to greater than the mark of the algoscopy for assessing marker levels
Marker levels in the test specimen of quasi-error, and preferably marker levels is (such as normal from having in control sample
The experimenter of glucose tolerance, there is glucemia is impaired on an empty stomach experimenter, there is the experimenter of impaired glucose tolerance, in the past
The average level of mark in the sample of the experimenter of type 2 diabetes mellitus and/or several control sample it has been diagnosed as) in 18 months
At least 1.5 times and more preferably 3,4,5,6,7,8,9 or 10 or more times.
" the more low-level " or " level reduction " of mark is meant less than the mark of the algoscopy for assessing marker levels
The level of the mark in the test specimen of quasi-error, and preferably in control sample marker levels (such as from just having
The experimenter of often glucose tolerance, there is glucemia is impaired on an empty stomach experimenter, there is the experimenter of impaired glucose tolerance, in mistake
The average water of mark in the sample of the experimenter of type 2 diabetes mellitus and/or several control sample it has been diagnosed as in going 18 months
Flat) at most 1/2 and more preferably 1/3,1/4,1/5,1/6,1/7,1/8,1/9 or 1/10 or lower.
Term " known standard level " or " control level " refer to the generally acknowledged or predetermined level of mark, and it is used for
Relatively from the level of the mark in the sample of experimenter.In one embodiment, the control level of mark is based on coming
The level of the mark in the sample of experimenter with NGT.In another embodiment, mark
Control level is based on the level from the mark in the sample with one or more experimenters that glucemia is impaired on an empty stomach.Separately
In one embodiment, the control level of mark is based on the mark in the sample from the experimenter with impaired glucose tolerance
The level of thing.In another embodiment, the control level of mark has been diagnosed as 2 types based in carrying out the comfortable past 18 months
The level of the mark in the sample of the experimenter of diabetes.In one embodiment, from the mark in the sample of experimenter
The control level of will thing be before in the sample of experimenter the level of fixed mark.
In still another embodiment, the control level of mark based on give impaired to empty stomach glucemia, glucose is resistance to
Measure the level from the mark in the sample of experimenter before the impaired and/or therapy of type 2 diabetes mellitus.In another embodiment
In, the control level of mark is based on from having of not contacting with test compound, on an empty stomach glucemia is impaired, glucose tolerance is subject to
The level of the mark in the sample of the experimenter of damage and/or type 2 diabetes mellitus.In another embodiment, the comparison of mark
Level water based on the mark in the sample from the experimenter with NGT contacted with test compound
Flat.In one embodiment, the control level of mark is based on impaired, the impaired glucose tolerance and/or 2 from empty stomach glucemia
The expression of the mark in the sample of the animal model of patients with type Ⅰ DM；It is derived from that on an empty stomach glucemia is impaired, impaired glucose tolerance
And/or the expression of the mark in the sample of the cell of the animal model of type 2 diabetes mellitus or cell line.
In other embodiments, " comparison " level of mark can be by measuring available from suspecting sugar on an empty stomach in experimenter
Blood is impaired, impaired glucose tolerance and/or type 2 diabetes mellitus outbreak before experimenter Samples subjects, available from achieve being subject to
The level of the mark of examination person's sample etc. determines.
As used herein, term " patient " or " experimenter " refer to people and non-human animal, such as veterinary patient.Term
" non-human animal " includes all vertebratess, such as mammal and nonmammalian, such as non-human primate, mice,
Rabbit, sheep, Canis familiaris L., cat, horse, milch cow, chicken, Amphibian and reptile.In one embodiment, described experimenter is people.
In some embodiments, the Body Mass Index (BMI) of experimenter is less than about 40kg/m2(e.g., from about 40,39,38,
37,36,35,34,33,32,31,30,29,28,27,26,25,24,23,22,21,20,19 or about 18kg/m2).Real at other
Executing in scheme, the Body Mass Index (BMI) of experimenter is greater than about 40kg/m2(e.g., from about 41,42,43,44,45,46,47,48,
49、50、51、52、53、54、55、56、57、58、59、60、61、62、63、64、65、66、67、68、69、70、71、72、73、
74,75,76,77,78,79 or about 80kg/m2)。
Term used herein " sample " refers to the gleanings of the similar cell or tissue separated from experimenter, and is being subject to
Tissue, cell and the fluid existed in examination person.Term " sample " include any body fluid from experimenter (such as blood, lymph,
Gynecological's fluid, capsule liquid, urine, tear and the fluid collected by bronchial lavage and/or peritoneum flushing) or cell.At one
In embodiment, take out described tissue or cell from experimenter.In another embodiment, described or cell is subject to described in being present in
In examination person.Other Samples subjects includes tear, serum, cerebrospinal fluid, feces, expectorant and cell extract.An embodiment
In, biological sample comprises to come the protein molecular of self-test experimenter.In another embodiment, biological sample can comprise to come
The mRNA molecule of self-test experimenter or carry out the genomic DNA molecule of self-test experimenter.
Term " measures " and means to include detecting mark in the presence or absence of mark in sample, quantitative determination sample
Amount and/or determine the method for type of biomarker.Measurement by methods known in the art and can be retouched the most further
Those methods stated realize.
Test kit is at least one reagent such as probe, primer or the antibody of the mark comprising the specific detection present invention
Any goods (such as packaging or container), described goods promote as a unit of the method for carrying out the present invention,
Distribution or sale.In certain embodiments, test kit can include substrate, such as, comprise one or more marks of the present invention
Capture agent and/or the substrate of capture agent that is combined with one or more marks of the present invention.In some embodiments
In, such test kit comprises the operation instructions of the level for using mass spectroscopy mark.
The mark of the present invention
One aspect of the present invention, it is provided that one is used for determining whether experimenter has or by Developing Grape impaired glucose tolerance
Maybe by the mark of development type 2 diabetes mellitus, comprising: cystine, glucose, sorbose, agedoite, lysine, different bright ammonia
Acid, threonine, fumaric acid, hydroxyurea, xylose, tyrosine, leucine, aspartic acid, glyceryl monostearate, linoleic acid, sweet
One or more in oil, β-mannose monoglyceride, D-talose.
Cystine (cystine): another name cystine；3,3'-dithio dipropyl propylhomoserin.Cystine is Branchamin
Thing, can promote cell Redox function, makes liver function vigorous, and can neutralize a toxin, promotes leucocyte hyperplasia, stop cause of disease
Bacterium grows.
Glucose (glucose): chemical formula C6H12O6, it is also called glucose, corn sugar, referred to as glucose.Fructus Vitis viniferae
Sugar has critical role in field of biology, is energy source and the metabolism intermediate product of living cells, and i.e. biological is main
Energy supply material.The function of detoxification of liver can be promoted, effect protected to liver.
Sorbose (sorbose): molecular formula C6H12O, belongs to one of ketose in monosaccharide, white crystal or crystalline powder, uses
Metabolism in animal and microorganism is studied.
Agedoite (asparagine): L-asparagine, linear formula H2NCOCH2CH(NH2)COOH。
Lysine (lysine): C6H14N2O2, lysine is one of essential amino acid, can promote human development, enhancing
Immunologic function, and it is improved the effect of central nervous tissue function.Lysine function in vivo has: participate in body protein such as skeleton
The synthesis of flesh, enzyme and polypeptide hormone；It is one of ketogenic amino acid, when lacking available carbohydrate, may participate in generation
The metabolism (under fasted conditions, be one of important energy source) of ketoboidies and glucose；Maintain internal acid-base balance；As conjunction
Become the precursor of carnitine, participate in lipid metabolism；It addition, lysine can also improve body opposing stress ability.
Isoleucine (Isoleucine): C6H13NO2, the effect of isoleucine includes closing together with leucine and valine
Make to repair muscle, control blood glucose, and provide energy to bodily tissue.
Threonine (threonine): C4H9NO3, it is a kind of essential amino acid, threonine metabolic pathway in body
Different with other aminoacid, it is unique without dehydrogenase effect and transamination, but by threonine dehydratase (TDH)
It is catalyzed with threonine dehydratase (TDG) and aldolase and is changed into the aminoacid of other materials.Approach mainly has 3: pass through aldehyde
Contracting enzymes metabolism is glycine and acetaldehyde；It is alanine, glycine, acetyl-COA by TDG metabolism；It is propanoic acid by TDH metabolism
And butyrine.
Fumaric acid (fumaric acid): C4H4O4, fumarate discharges free radical in removing inflammatory process, thus protects
Protect nerve and glial cell.
Xylose (xylose): xylose is a component of xylan, and xylan is widely present in plant.Xylose there is also
In animal heparin, chrondroitin and glycoprotein, it is sugar chain and the connection unit of serine (or threonine) in some glycoprotein.
Tyrosine (tyrosine): C9H11NO3。
Leucine (leucine): C6H13NO2, leucic effect includes cooperating to repair together with isoleucine and valine
Multiple muscle, controls blood glucose, and provides energy to bodily tissue.
Aspartic acid (aspartic acid): also known as aspartic acid, is a kind of a-amino acid, and aspartic acid is generally deposited
It is in biosynthesis effect.It is the aminoacid such as lysine, threonine, isoleucine, methionine and purine in organism, phonetic
The synthesis precursor of pyridine base.
Linoleic acid (linoleic acid): C18H32O2。
Glycerol (glycerol), β-mannose monoglyceride (beta-Mannosylglycerate), D-talose (D-
Talose)。
In certain aspects of the invention, single mark can be used for the method and composition of the present invention.Such as, a reality
Executing in scheme, the mark for the method and composition of the present invention is cystine.In one embodiment, mark is Portugal
Grape sugar.In one embodiment, mark is sorbose.In one embodiment, mark is agedoite.At one
In embodiment, mark is lysine.In one embodiment, mark is isoleucine.In one embodiment,
Mark is glyceryl monostearate.In one embodiment, mark is linoleic acid.In one embodiment, mark
Thing is glycerol.In one embodiment, mark is β-mannose monoglyceride.In one embodiment, mark is
D-talose.
In the other side of the present invention, exceed a kind of mark, such as multiple markers, such as 2,3,4,5,6,7,8,9,
10,11 or more kinds of mark, can be used for the method and composition of the present invention.Such as, in one embodiment, for this
The mark of bright method and composition includes cystine and glucose.In one embodiment, mark includes cystine
And sorbose.In one embodiment, mark includes cystine and agedoite.In one embodiment, mark
Including cystine and lysine.In one embodiment, mark includes cystine and isoleucine.An embodiment
In, mark includes cystine and threonine.In one embodiment, mark includes cystine and fumaric acid.At one
In embodiment, mark includes cystine and hydroxyurea.In one embodiment, mark includes cystine and xylose.
In one embodiment, mark includes cystine and tyrosine.In one embodiment, mark include cystine and
Leucine.In one embodiment, mark includes cystine and aspartic acid.In one embodiment, mark bag
Include cystine and glyceryl monostearate.In one embodiment, mark includes cystine and linoleic acid.An enforcement
In scheme, mark includes cystine and glycerol.In one embodiment, mark includes cystine and β-mannose glycerol
Acid esters.In one embodiment, mark includes cystine and D-talose.In one embodiment, mark includes
Glucose and sorbose.In one embodiment, mark includes glucose and agedoite.In one embodiment,
Mark includes glucose and lysine.In one embodiment, mark includes glucose and isoleucine.A reality
Executing in scheme, mark includes glucose and threonine.In one embodiment, mark includes glucose and fumaric acid.
In one embodiment, mark includes glucose and hydroxyurea.In one embodiment, mark include glucose and
Xylose.In one embodiment, mark includes glucose and tyrosine.In one embodiment, mark includes Portugal
Grape sugar and leucine.In one embodiment, mark includes glucose and aspartic acid.In one embodiment, mark
Will thing includes glucose and glyceryl monostearate.In one embodiment, mark includes glucose and linoleic acid.One
In individual embodiment, mark includes glucose and glycerol.In one embodiment, mark includes glucose and β-manna
Sugar monoglyceride.In one embodiment, mark includes glucose and D-talose.
In one embodiment, mark includes sorbose and agedoite.In one embodiment, mark bag
Include sorbose and lysine.In one embodiment, mark includes sorbose and isoleucine.An embodiment
In, mark includes sorbose and threonine.In one embodiment, mark includes sorbose and fumaric acid.At one
In embodiment, mark includes sorbose and hydroxyurea.In one embodiment, mark includes sorbose and xylose.
In one embodiment, mark includes sorbose and tyrosine.In one embodiment, mark include sorbose and
Leucine.In one embodiment, mark includes sorbose and aspartic acid.In one embodiment, mark bag
Include sorbose and glyceryl monostearate.In one embodiment, mark includes sorbose and linoleic acid.An enforcement
In scheme, mark includes sorbose and glycerol.In one embodiment, mark includes sorbose and β-mannose glycerol
Acid esters.In one embodiment, mark includes sorbose and D-talose.
In one embodiment, mark includes agedoite and lysine.In one embodiment, mark bag
Include agedoite and isoleucine.In one embodiment, mark includes agedoite and threonine.An embodiment party
In case, mark includes agedoite and fumaric acid.In one embodiment, mark includes agedoite and hydroxyurea.
In one embodiment, mark includes agedoite and xylose.In one embodiment, mark includes agedoite
And tyrosine.In one embodiment, mark includes agedoite and leucine.In one embodiment, mark
Including agedoite and aspartic acid.In one embodiment, mark includes agedoite and glyceryl monostearate.?
In one embodiment, mark includes agedoite and linoleic acid.In one embodiment, mark includes agedoite
And glycerol.In one embodiment, mark includes agedoite and β-mannose monoglyceride.An embodiment
In, mark includes agedoite and D-talose.
In one embodiment, mark includes cystine, glucose and sorbose.In one embodiment, mark
Will thing includes cystine, glucose and agedoite.In one embodiment, mark includes cystine, glucose and relies
Propylhomoserin.In one embodiment, mark includes cystine, glucose and isoleucine.In one embodiment, mark
Thing includes cystine, glucose and fumaric acid.In one embodiment, mark include, D-talose, β-mannose glycerol
Acid esters and sorbose.In one embodiment, mark includes cystine, β-mannose monoglyceride and linoleic acid.One
In individual embodiment, mark includes β-mannose monoglyceride, glyceryl monostearate and sorbose.An embodiment
In, mark includes β-mannose monoglyceride, D-talose and glyceryl monostearate.In one embodiment, mark
Thing includes β-mannose monoglyceride, glucose and leucine.In one embodiment, mark includes β-mannose glycerol
Acid esters, glycerol and sorbose.In one embodiment, mark includes β-mannose monoglyceride, D-talose and Guang ammonia
Acid.In one embodiment, mark includes cystine, glucose, sorbose and agedoite.An embodiment
In, mark includes cystine, glucose, sorbose and lysine.In one embodiment, mark include cystine,
Glucose, sorbose and isoleucine.In one embodiment, mark includes cystine, glucose, sorbose and Soviet Union's ammonia
Acid.In one embodiment, mark includes cystine, glucose, sorbose and fumaric acid.In one embodiment,
Mark includes cystine, glucose, sorbose, agedoite and lysine.In one embodiment, mark includes Guang
Propylhomoserin, glucose, sorbose, agedoite and isoleucine.In one embodiment, mark includes cystine, Fructus Vitis viniferae
Sugar, sorbose, agedoite and threonine.In one embodiment, mark includes cystine, glucose, sorbose, sky
Winter amide, isoleucine and lysine.In one embodiment, mark includes cystine, glucose, sorbose, Radix Asparagi
Amide, isoleucine, threonine and lysine.In one embodiment, mark include cystine, glucose, sorbose,
Agedoite, isoleucine, threonine, fumaric acid and lysine.In one embodiment, mark includes cystine, Portugal
Grape sugar, sorbose, agedoite, isoleucine, threonine, fumaric acid, hydroxyurea and lysine.In one embodiment,
Mark include cystine, glucose, sorbose, agedoite, isoleucine, threonine, fumaric acid, hydroxyurea, xylose and
Lysine.In one embodiment, mark includes cystine, glucose, sorbose, agedoite, isoleucine, Soviet Union's ammonia
Acid, fumaric acid, hydroxyurea, tyrosine and lysine.In one embodiment, mark includes cystine, glucose, Pyrusussuriensis
Sugar, agedoite, isoleucine, threonine, fumaric acid, hydroxyurea, tyrosine, leucine and lysine.An embodiment party
In case, mark includes cystine, glucose, sorbose, agedoite, isoleucine, threonine, fumaric acid, hydroxyurea, cheese
Propylhomoserin, leucine, aspartic acid and lysine.In one embodiment, mark include cystine, glucose, sorbose,
Agedoite, isoleucine, threonine, fumaric acid, hydroxyurea, tyrosine, leucine, aspartic acid, glyceryl monostearate
And lysine.In one embodiment, mark includes cystine, glucose, sorbose, agedoite, isoleucine, Soviet Union
Propylhomoserin, fumaric acid, hydroxyurea, tyrosine, leucine, aspartic acid, glyceryl monostearate, linoleic acid and lysine.One
In individual embodiment, mark include cystine, glucose, sorbose, agedoite, isoleucine, threonine, fumaric acid,
Hydroxyurea, tyrosine, leucine, aspartic acid, glyceryl monostearate, linoleic acid, glycerol and lysine.
In one embodiment, mark includes cystine, glucose, sorbose, agedoite, isoleucine, Soviet Union
Propylhomoserin, fumaric acid, hydroxyurea, tyrosine, leucine, aspartic acid, glyceryl monostearate, linoleic acid, glycerol, β-mannose
Monoglyceride and lysine.In one embodiment, mark includes cystine, glucose, sorbose, isoleucine, Soviet Union
Propylhomoserin, fumaric acid, hydroxyurea, tyrosine, leucine, aspartic acid, glyceryl monostearate, linoleic acid, glycerol, β-mannose
Monoglyceride, D-talose and lysine.
In one embodiment, mark includes cystine, glucose, sorbose, agedoite, isoleucine, Soviet Union
Propylhomoserin, fumaric acid, hydroxyurea, tyrosine, leucine, aspartic acid, glyceryl monostearate, linoleic acid, glycerol, β-mannose
Monoglyceride and lysine.In one embodiment, mark includes cystine, glucose, sorbose, agedoite, different
Leucine, threonine, fumaric acid, hydroxyurea, tyrosine, leucine, aspartic acid, glyceryl monostearate, linoleic acid, sweet
Oil, β-mannose monoglyceride, D-talose and lysine.
One aspect of the present invention provides the method obtaining described mark, and its concrete steps are:
1, obtain Samples subjects, and carry out pre-treatment；
2, the Samples subjects of pre-treatment is carried out GC-MS or LC-MS to detect, collect data；
3, carry out data analysis, obtain feasible mark.
In above-mentioned steps, described Samples subjects can be solid, liquid, it is also possible to is cell, microorganism etc..
As a kind of embodiment, when described Samples subjects is solid, the method for described concrete pre-treatment is: will
100mg solid is dissolved in 400 μ L and extracts reagent, adds steel ball and is ground, and the condition of described grinding is 30Hz, 5 minutes；Then enter
Row is centrifugal, takes supernatant.
To carry out GC-MS test, can be concentrated dry.The step of described concentrate drying is will to add in supernatant
The methoxamine of 20mg/ml, is dried 20min at 80 DEG C, is subsequently adding under reagent BSTFA after performing the derivatization process 1 hour, enters
Row test.
To carry out LC-MS test, then only supernatant need to be carried out concentrate drying, add 100 μ L acetonitriles and redissolve, warp
Filtering plug, can directly carry out LC-MS test.
As a kind of embodiment, when described Samples subjects is liquid, such as serum, urine etc..Described concrete front place
The method of reason is: 100 μ L liquid add in 350 μ L methanol, carries out mixing of mediating, high speed centrifugation, takes supernatant, test.
To carry out GC-MS test, can be concentrated dry.The step of described concentrate drying is will to add in supernatant
The methoxamine of 20mg/ml, is dried 20min at 80 DEG C, is subsequently adding under reagent BSTFA after performing the derivatization process 1 hour, enters
Row test.
To carry out LC-MS test, then only supernatant need to be carried out concentrate drying, add 100 μ L acetonitriles and redissolve, warp
Filtering plug, can directly carry out LC-MS test.
As a kind of embodiment, when described Samples subjects is for microorganism, described processing mode and the process side of cell
Formula is the same.
The method of the present invention
In some aspects, the present invention provides diagnostic method.Such as, in one aspect, the present invention is provided to determine tested
Whether person has the method for impaired glucose tolerance.One that described method includes measuring the present invention in Samples subjects or
The level of multiple markers and the level of one or more marks described in control sample.A kind of with control sample or
The level of multiple markers is compared, the level difference (the most higher or lower) of one or more marks in Samples subjects
Described experimenter is indicated to have impaired glucose tolerance.
On the other hand, the present invention is provided to determine the method whether experimenter has type 2 diabetes mellitus.Described method bag
Include level and the described one in control sample of one or more marks of the present invention measured in Samples subjects
Or the level of multiple markers.Compared with the level of one or more marks in control sample, in Samples subjects
Plant or the level difference (the most higher or lower) of multiple markers indicates described experimenter to have type 2 diabetes mellitus.
The present invention also provides for method of prognosis.Such as, in one aspect, the present invention is provided to determine whether experimenter will send out
The method of exhibition impaired glucose tolerance.Described method includes one or more marks measuring the present invention in Samples subjects
The level of thing and the level of one or more marks described in control sample.With one or more marks in control sample
The level of will thing is compared, and the level difference (the most higher or lower) of one or more marks in Samples subjects indicates institute
State experimenter by Developing Grape impaired glucose tolerance.
On the other hand, the present invention is provided to determine that whether experimenter is by the method for development type 2 diabetes mellitus.Described method
Including the level of one or more marks of the present invention measured in Samples subjects and described in control sample
Plant or the level of multiple markers.Compared with the level of one or more marks in control sample, in Samples subjects
Plant or the level difference (the most higher or lower) of multiple markers indicates described experimenter will develop type 2 diabetes mellitus.
Multiple complications is relevant to impaired glucose tolerance and/or type 2 diabetes mellitus, especially with long-term glucose tolerance
Impaired and/or type 2 diabetes mellitus is correlated with.Such as, such experimenter has the cardiovascular disease of 2-4 times of risk and (includes ischemic
Heart disease and stroke), lower extremity amputation increases by 20 times and admission rate and increases.Type 2 diabetes mellitus is also atraumatic blinding and nephropathy (bag
Include renal failure) main reason, and with cognitive dysfunction and dementia (by lysis such as Alzheimer and blood
Pipe is dull-witted) risk increase relevant.Other complication includes such as neuropathy, acanthosis nigricans, sexual dysfunction and often feels
Dye.
Newly it is being diagnosed as the experimenter of type 2 diabetes mellitus because the mark of the present invention has shown and is having been determined as 2 type glycosurias
Difference table in sick those experimenters (such as there are those experimenters of long-term impaired glucose tolerance and/or type 2 diabetes mellitus)
Reaching, therefore the present invention also provides for for determining that whether experimenter is by the method for development type 2 diabetes mellitus related complication.Described method
Including the level of one or more marks of the present invention measured in Samples subjects and described in control sample
Plant or the level of multiple markers.Compared with the level of one or more marks in control sample, in Samples subjects
The level difference (the most higher or lower) of one or more marks indicates described experimenter will respond diabetotherapy.
On the other hand, the present invention is provided to determine the experimenter with impaired glucose tolerance and/or type 2 diabetes mellitus
The method that whether therapeutic scheme will be responded.Described method includes the one or many measuring the present invention in Samples subjects
Plant the level of mark and the level of one or more marks described in control sample.With the one in control sample or
The level of multiple markers is compared, the level difference (the most higher or lower) of one or more marks in Samples subjects
Described experimenter is indicated will treatment to be responded.
Multiple diabetotherapy known in the art, including such as insulin sensitizers, such as (such as diformazan is double for biguanides
Guanidine) and thiazolidinediones (such as rosiglitazone, pioglitazone, troglitazone)；Succagoga, such as sulphanylureas (such as lattice
Arrange this urea, glipizide, glimepiride, tolbutamide, acetohexamide, tolazamide, chlorpropamide, gliclazide, lattice row
Quinoline ketone), non-sulphanylureas succagoga such as meglitinide derivant (such as repaglinide, Nateglinide)；DPP IV
Inhibitor (such as sitagliptin, BMS-477118, BI 1356, vildagliptin, Egelieting)；Alpha-glucosidase inhibitor is (such as
Acarbose, miglitol, voglibose)；Diabetes-associated peptide analogies (such as pramlintide acetate)；Intestinal blood sugar lowering
Mimetics (such as Exenatide, Arg34Lys26-(N-EPSILON-(N-ALPHA-Palmitoyl-L-GAMMA-glutamyl))-GLP-1[7-37], taspoglutide)；Insulin and its analog (such as quick acting, slowly rise
Effect and medium onset)；Bile acid multivalent chelator (such as colesevelam)；With dopamine agonist (such as bromocriptine), individually
Or be applied in combination.
In certain embodiments of the invention, described treatment includes insulin sensitizers.In another embodiment, institute
State treatment and include insulin sensitizers and succagoga.In still another embodiment, described treatment include insulin sensitizers,
Succagoga and insulin.
The method of the present invention can be with technical staff for diagnosing, predict and/or monitor the impaired glucose tolerance of experimenter
And/or type 2 diabetes mellitus and/or type 2 diabetes mellitus complication and/or any other method of the reaction for the treatment of is implemented together.Example
As, the method for the present invention can be entered together with any clinical measurement of glucose tolerance known in the art, obesity and/or diabetes
OK, described measurement include the serology of other molecular marker, cytology and/or detection (with quantitatively, if appropriate).
In any method (and test kit) of the present invention, available from the level of the mark of the present invention in the sample of experimenter
Can be measured by any one in various widely-known techniques and method, the mark of the present invention in sample is converted by it
One-tenth can detect and quantitative part.The limiting examples of such method includes using for the immunology side detecting protein
Method, method of purifying protein, protein function or activation measurement, nucleic acid hybridization, nucleic acid reverse-transcription method and nucleic acid expand
Increasing method, immunoblotting, western blotting, RNA blotting, electron microscopy, mass spectrography (such as MALDI-TOF and
SELDI-TOF), immunoprecipitation, immunofluorescence, immunohistochemistry, enzyme-linked immunosorbent assay (ELISA) (such as expand
ELISA), quantitative determination process based on blood (such as serum ELISA), quantitative determination process based on urine, flow cytometry,
DNA hybridization, array analysis etc. and a combination thereof or sub-combination analyze sample.
Those of ordinary skill it will be readily understood that, this area has been set up for detecting this on nucleic acid or protein level
Any technological means of the level of bright mark is basically available for measuring the water of the mark of invention as described herein
Flat.
In one embodiment, the level of the mark of the present invention uses nucleic probe to measure.Art used herein
Language " probe " refers to any molecule of the special sign thing of the selective binding present invention.Probe can be by those skilled in the art
Synthesis, or derive from suitable Biological preparation.Probe can be specially designed for labelling.Can be used as the example of the molecule of probe
Include but not limited to RNA, DNA, protein, antibody and organic molecule.
In one embodiment of the invention, microarray is used to detect the level of mark of the present invention.Because no
With the reproducibility between experiment, microarray is highly suitable for this purpose especially.DNA microarray provides one for simultaneously
The method measuring the level of lots of genes.Each array is made up of the capture probe of the reproducible form being attached to solid support.
RNA or DNA of labelling and the complementary probe hybridization on array, then pass through laser scanning inspection.Each probe on array miscellaneous
Hand over intensity after measured and change into the quantitative values representing relative gene expression level.
Following methods is included, such as electricity for detecting other known method of the mark of the present invention on protein level
Swimming, capillary electrophoresis, high performance liquid chromatography, thin layer chromatography, super diffusion chromatograph etc., or various immunological method such as fluid or solidifying
Glue precipitation, immunodiffusion, immunoelectrophoresis, radioimmunoassay, enzyme-linked immunosorbent assay, immunofluorescence assay
And western blotting.
In one embodiment of the invention, proteomic approach, such as mass spectrography are used.Mass spectrography is a kind of analysis
Technology, with generation charged molecule (or its fragment) and measures its mass-charge ratio including by compound ionization.At typical mass spectrum
In program, obtain sample from experimenter, be loaded on mass spectrum, and its component (mark of the such as present invention) is by different sides
Method ionization (such as by with beam bombardment they), results in charged particle (ion).Then electromagnetic field is passed through when ion
Time, according to the mass-charge ratio of the motion calculation granule of ion.
Such as, mastrix-assisted laser desorption ionization time of flight mass spectrography or the surface-enhanced laser desorbing/ionization of Matrix-assisted is flown
Time mass spectrum method, it includes that application biological sample such as serum to protein binding chip can be used for measuring the mark of the present invention
Level.
Additionally, the vivo techniques of the level for measuring mark of the present invention includes the labelling for mark of the present invention
Antibody is incorporated in experimenter, and the mark of the described antibodies present invention also converts it into detectable molecule.As above institute
Stating, the existence of the detected mark of the present invention in experimenter, level or even position can be detected by standard imaging techniques
And measure.
Generally speaking, the preferably level of the mark of the present invention in Samples subjects and the mark of the present invention in control sample
Difference between the amount of thing is the biggest.Although this difference may be with the detection limit of the method for measuring marker levels
The least, but preferred described difference is the standard error of appraisal procedure at least above the standard error of appraisal procedure, preferably difference
At least 2,3,4,5,6,7,8,9,10,15,20,25,100,500,1000 times or bigger.
For the method monitoring therapeutic efficiency
The present invention also provides for can be used for suppressing impaired glucose tolerance and/or type 2 diabetes mellitus in experimenter for monitoring
Development；Reduce or the NGT that slows down is impaired to empty stomach glucemia, to impaired glucose tolerance and/or to diabetes
Progress；And/or reduce or the therapy of development of the suppression complication relevant to described disease or therapeutic scheme or any other are controlled
The method of effect for the treatment of method.In these methods, assessment is a pair sample (the first sample and process without therapeutic scheme
At least one of second sample of therapeutic scheme) in the level of one or more marks of the present invention.Relative to second
Sample, in the first sample, the regulation of the expression of one or more marks shows that described therapy effectively suppresses in experimenter
Impaired glucose tolerance and/or the development of type 2 diabetes mellitus；Reduce or the NGT that slows down is impaired to empty stomach glucemia, extremely
Impaired glucose tolerance and/or the progress to diabetes；And/or reduce or the suppression complication relevant to described disease
Exhibition.
The method identifying type 2 diabetes mellitus biomarker
The present invention further provides the method for identifying type 2 diabetes mellitus biomarker, described biomarker is used as
The therapeutic efficiency (treatment diagnosis) of such as disease (prognosis and diagnosis), medicine and the mark of drug toxicity.Such as, as above institute
Stating, the mark that mark as herein described and the method using biomarker to find are identified can be used for such as determining experimenter
Whether have or by Developing Grape impaired glucose tolerance；Determine experimenter whether to have maybe and will develop type 2 diabetes mellitus；Determine have 2
Whether the experimenter of patients with type Ⅰ DM will respond diabetotherapy；Monitoring is for suppressing impaired glucose tolerance in experimenter
And/or the development of type 2 diabetes mellitus, reduce or the NGT that slows down is impaired to empty stomach glucemia, to impaired glucose tolerance
And/or effect of therapy of the development to the progress of diabetes and/or reduction or the suppression complication relevant to described disease；Mirror
Joint (such as reduce or increase) of setting the tone is used as the expression of the mark of the present invention of such as therapy and/or the screening of the molecule of activity
In algoscopy.
Type 2 diabetes mellitus mark is identified also by following: measure before providing at least some of therapy to experimenter
Available from the protein level in first sample of the experimenter with type 2 diabetes mellitus, and measure at least one of the described therapy of offer
Available from the protein level in second sample of described experimenter after Fen.Relative to the first sample, protein expression water in the second sample
Flat difference, such as statistical significant level, identify that described albumen is type 2 diabetes mellitus mark.
The test kit of the present invention
The present invention also provides for for determining whether experimenter has or by Developing Grape impaired glucose tolerance and/or experimenter be
No have maybe will development type 2 diabetes mellitus test kit.Also provide for determining that experimenter whether will development type 2 diabetes mellitus complication, really
Whether constant current modulation method will effectively be treated the test kit of the experimenter with impaired glucose tolerance and/or type 2 diabetes mellitus and is used for supervising
Survey the test kit of effect of therapy.
These test kits include that the instrument measuring the level of one or more marks of the present invention and test kit use and say
Bright book.
The test kit of the present invention can optionally comprise the other component for carrying out the inventive method.Such as, test kit can wrap
Containing for obtaining the reagent of biological sample, control sample, one or more sample compartment, Remedies for diabetes from experimenter, retouch
State guiding material and the non-tissue specific control/standard substance carrying out the inventive method.
For measure the reagent of the level of one or more marks can include such as buffer or for evaluate one or
Other reagent of the algoscopy of the level of multiple markers.Description can be such as be evaluated the present invention one or
The description of the printing of the algoscopy of the level of multiple markers.
Can include can be used for obtaining the one of fluid or tissue from experimenter for separating the reagent of biological sample from experimenter
Plant or plurality of reagents, such as obtaining the instrument of saliva or blood.
The test kit of the present invention can farther include the reagent for cultivating the sample available from experimenter.
Preferably, test kit is designed for human experimenter.
The present invention is described further by following example, and embodiment should not be construed as restriction further.The application is complete
All lists of references, patent and the content of disclosed patent application cited in literary composition, and accompanying drawing, explicitly by quoting with it
It is incorporated integrally into herein.
Equivalent
In describing exemplary embodiment, the reason for clarification uses specific term.For describe purpose, each specific
Term mean at least to include all technology and functional equivalent, it operates the purpose realizing being similar in a similar fashion.This
Outward, the most concrete exemplary embodiment includes in some examples of multiple factor of system or method step, those key elements
Or step can be replaced single key element or step.Equally, single key element or step can be replaced and want for the multiple of identical purpose
Element or step.Additionally, specify the parameter of various character of exemplary embodiment herein in the case of, those parameters can be upwards
Or adjust downwards 1/20,1/10,1/5,1/3 etc., or adjusted, except as otherwise noted by its approximation that rounds up.Additionally, to the greatest extent
Pipe exemplary embodiment is shown and described by reference to its specific embodiment, but those of ordinary skill in the art will
Understand, in the case of without departing substantially from the scope of the present invention, it can be carried out the various replacements in form and details and change.The most another
Outward, other side, function and advantage are also within the scope of the invention.
There is provided herein the flow chart of exemplary for illustrative purposes, and be nonrestrictive method example.Ability
Territory skilled artisan will realize that, the method for exemplary can include that those than illustrating in exemplary flow chart are more or less
Step, and the step in exemplary flow chart can carry out from shown different order.
Packet: be divided into the complication group that normal group, type 2 diabetes mellitus group, prediabetes group, type 2 diabetes mellitus are relevant.
Choose 1250 experimenters, wherein normal healthy people 320 people；Type 2 diabetes mellitus 300 people；Prediabetes 330 people；2
Complication 300 people that patients with type Ⅰ DM is relevant.
One, mark is obtained
Step: 1, obtain experimenter's serum, and carry out pre-treatment: 100 μ L liquid add in 350 μ L methanol, mediate
Mixing, high speed centrifugation, take supernatant, carry out concentrate drying, add 100 μ L acetonitriles and redissolve, through filtering plug, can directly carry out
GC-MS tests；Collecting GC-MS data, the mode carrying out principal component analysis counts suitable mark.Principal component analysis figure is such as
Shown in Figure 10.
Shown in the principal component analysis figure of Figure 10, normal group can substantially be distinguished substantially with diabetic groups sample, logical
Later continuous discriminant analysis studies its difference further.
Find, after using GC-MS quantitative verification, the complication that type 2 diabetes mellitus group, prediabetes group, type 2 diabetes mellitus are relevant
Group, compared with normal person's group, special discrepant expression material has: cystine, glucose, sorbose, agedoite, bad ammonia
Acid, isoleucine, threonine, fumaric acid, hydroxyurea, xylose, tyrosine, leucine, aspartic acid, glyceryl monostearate,
Linoleic acid, glycerol, β-mannose monoglyceride, D-talose, result is as shown in table 1:
The table 1 diabetic groups contrast special discrepant expression material of normal group
Wherein, fold change value refers to that the average of diabetic groups something content organizes the equal of this content of material divided by normal person
Value；
Variable importance (VIP): the something difference between the two groups contribution degree to group difference between group, it is considered that
The material contribution degree of VIP > 1 is notable, and VIP is the biggest, and contribution degree is the biggest；
Similarity: when material is qualitative, the marking value after inspection mass spectrum is compared with the mass spectrum in standard spectrum storehouse in fact,
Full marks 1000 points, similar with standard mass spectrum closer to 1000 bright real inspection mass spectruies of defending oneself, it is qualitative the most accurate, typically to be equivalent to
Think similarity > 800 for qualitative accurately.
Embodiment is as shown in table 2:
Table 2 detailed description of the invention
Wherein, numbering " 2-3 " represents and diagnoses 2 type sugar with the mark of numbered 2 and the mark combinatorial association of numbered 3
Urine disease group and normal person's group, namely diagnose with glucose and glycerol for mark combinatorial association.Remaining numbering is by that analogy.
Comparative example 1: deubiquitinating enzymes and neurofascin
With deubiquitinating enzymes and neurofascin Combining diagnosis type 2 diabetes mellitus group and normal person's group.
Shown in result such as Fig. 1 ROC curve (Receiver operating curve), it is known that AUC (area under ROC curve) is
0.7436, diagnostic sensitivity is 49.81%, and specificity is 15.4%.
When data above is it can be seen that mark is applied in combination, diagnostic sensitivity is 96.3%, and specificity is 0%.This
Bright selected mark has the strongest prospect of the application, is thus provided that the Advantageous Effects of the present invention.
Aforesaid example is merely illustrative, for explaining some features of the feature of the disclosure.Appended claim
It is intended to the widest scope that requirement it is contemplated that, and embodiments as presented herein is only according to all possible embodiment
The explanation of embodiment of selection of combination.Therefore, the purpose of applicant is that appended claim is not by the explanation present invention
The selectional restriction of example of feature.And the progress in science and technology will be formed due to language performance inaccurate reason and not
The possible equivalent or the son that are presently considered are replaced, and these changes also should be interpreted in the conceived case by appended
Claim covers.
Claims (10)
1. for determining whether experimenter has or by Developing Grape impaired glucose tolerance maybe by the mark of development type 2 diabetes mellitus,
Comprising: one or more in glucose, β-mannose monoglyceride, D-talose.
2. it is used for determining whether experimenter has or development type 2 diabetes mellitus maybe maybe will be developed 2 by Developing Grape impaired glucose tolerance
The method of patients with type Ⅰ DM related complication, described method includes:
The level of one or more marks in claim 1 is measured in the sample of experimenter；
Water by the level of one or more marks in Samples subjects with one or more marks in control sample
Put down and compare, the one or many wherein compared with the level of one or more marks in control sample, in Samples subjects
The difference of the level planting mark indicates described experimenter to have or by Developing Grape impaired glucose tolerance.
3. for determining that there is the side whether experimenter of impaired glucose tolerance and/or type 2 diabetes mellitus will respond therapy
Method, described method includes
The level of one or more marks in claim 1 is measured in the sample of experimenter；
Water by the level of one or more marks in Samples subjects with one or more marks in control sample
Put down and compare, the one or many wherein compared with the level of one or more marks in control sample, in Samples subjects
The difference of the level planting mark indicates described experimenter will respond described therapy.
4. it is used for determining whether experimenter has or development type 2 diabetes mellitus maybe maybe will be developed 2 by Developing Grape impaired glucose tolerance
The test kit of patients with type Ⅰ DM complication, described test kit includes the one in claim 1 in Samples subjects or many
Plant the reagent of the level of mark and determine whether experimenter has or Developing Grape impaired glucose tolerance maybe will develop 2 type glycosurias
Sick maybe by the test kit operation instructions of development type 2 diabetes mellitus complication.
5. for determining the examination whether experimenter with impaired glucose tolerance and/or type 2 diabetes mellitus will respond to treatment
Agent box, described test kit includes the level for measuring one or more marks in claim 1 in Samples subjects
Reagent and determine the test kit operation instructions whether experimenter will respond to described treatment.
6. it is used for determining whether experimenter has or development type 2 diabetes mellitus maybe maybe will be developed 2 by Developing Grape impaired glucose tolerance
The method of patients with type Ⅰ DM related complication, described method includes
The level of the mensuration described cystine in Samples subjects；
By the level of the cystine in Samples subjects compared with the level of the cystine in control sample, wherein experimenter's sample
The level of the cystine in product, higher than the level of the cystine in control sample, indicates described experimenter to have or by Developing Grape
Impaired glucose tolerance.
7. for determining the side whether experimenter with impaired glucose tolerance and/or type 2 diabetes mellitus will respond to treatment
Method, described method includes
The level of mensuration cystine in Samples subjects；
By the level of the cystine in Samples subjects compared with the level of cystine in control sample, wherein Samples subjects
In the level of cystine higher than the level of the cystine in control sample, indicate described experimenter will described treatment be had instead
Should.
8. the method monitoring effect for the treatment of in there is the experimenter of impaired glucose tolerance and/or type 2 diabetes mellitus, described side
Method includes:
Give described in experimenter treatment at least some of before and after, measure the water of cystine in Samples subjects
Flat；With
By before giving described treatment from the cystine in Samples subjects level with giving described treatment at least
After a part, the level from the cystine in Samples subjects compares, wherein with giving the sample before described treatment
In the level of cystine compare, give described treatment at least some of after sample in the level of cystine reduce
Show that described treatment is responded by described experimenter, thus monitor effect of described treatment.
9. the method any one of claim 6-8, farther includes to measure in the claim 1 in Samples subjects
Plant or the level of multiple other mark.
10. the method described in claim 1, described mark also includes: cystine, sorbose, agedoite, lysine, different
Leucine, threonine, fumaric acid, hydroxyurea, xylose, tyrosine, leucine, aspartic acid, glyceryl monostearate, sub-oil
One or more in acid, glycerol.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610393254.3A CN106093430A (en) | 2016-06-06 | 2016-06-06 | Can be used for mark detecting diabetes and application thereof |
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| CN201610393254.3A CN106093430A (en) | 2016-06-06 | 2016-06-06 | Can be used for mark detecting diabetes and application thereof |
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Family
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| CN201610393254.3A Pending CN106093430A (en) | 2016-06-06 | 2016-06-06 | Can be used for mark detecting diabetes and application thereof |
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| CN (1) | CN106093430A (en) |
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| CN108152502A (en) * | 2017-11-23 | 2018-06-12 | 上海阿趣生物科技有限公司 | Composite marker object available for detecting diabetes early stage and application thereof |
| CN108196052A (en) * | 2017-11-23 | 2018-06-22 | 上海阿趣生物科技有限公司 | Composite marker object available for detecting the cancer of the esophagus and application thereof |
| CN112903840A (en) * | 2021-01-18 | 2021-06-04 | 江苏省中医院 | Application of serum metabolite in preparation of kit for diagnosing sjogren's syndrome |
| CN115023608A (en) * | 2021-11-30 | 2022-09-06 | 江苏品生医疗科技集团有限公司 | Marker for predicting possibility of diabetes of subject and application thereof |
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Application publication date: 20161109 |