CN101551396A - Chemiluminscence immunoassay kit of hepatitis E virus IgG antibody and preparation method thereof - Google Patents
Chemiluminscence immunoassay kit of hepatitis E virus IgG antibody and preparation method thereof Download PDFInfo
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- CN101551396A CN101551396A CNA2008101032659A CN200810103265A CN101551396A CN 101551396 A CN101551396 A CN 101551396A CN A2008101032659 A CNA2008101032659 A CN A2008101032659A CN 200810103265 A CN200810103265 A CN 200810103265A CN 101551396 A CN101551396 A CN 101551396A
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Abstract
The invention discloses a chemiluminscence immunoassay kit of a hepatitis E virus IgG antibody and a preparation method thereof. The kit comprises negative and positive reference substances of the hepatitis E virus IgG antibody, a coated solid phase carrier, an enzyme labeling combination, sample diluent, chemiluminescent substrate solution and concentrated cleaning solution. The preparation method of the kit comprises the following steps: the reference substances are prepared according to the negative and the positive serums of the hepatitis E virus IgG antibody; the solid phase carrier is coated by recombining the antigen of the hepatitis E virus; the special anti-human IgG antibody is labeled by enzyme; the sample diluent is prepared; the chemiluminescent substrate solution is prepared; the concentrated cleaning solution is prepared; all the components are subpackaged; and the components are assembled into finished products. The kit can provide the most direct clinical basis for the prevention, the diagnosis and the treatment of the hepatitis E.
Description
Technical field
The present invention relates to in-vitro diagnosis immuno analytical method field, the chemiluminescence enzyme immunoassay that belongs to Hepatitis E IgG antibody test in the serum (oar) is measured kit.Simultaneously, the invention provides a kind of hepatitis E virus IgG antibody chemical luminescence immune assay determination reagent kit and preparation method thereof.
Background technology
(hepatitis E virus is nonencapsulated single positive chain RNA virus HEV) to hepatitis E virus, propagates through fecal-oral route, acute Hepatitis E (the hepatitisE that causes, HE), show as acute Jaundice Jaundice more, easily develop into subacute hepatitis sequestrans or silt courage hepatitis.Be ascendant trend year by year in recent years.The patient is mainly the adult, and case fatality rate is higher, especially last 3 months gravid woman of pregnancy period ill after, case fatality rate can reach 10%～39%, harm is serious.Viral hepatitis type E at first finds that at the Indian subcontinent the Central Asia, Southeast Asia, Africa, the Indian subcontinent all have more pandemic report.Most eruption and prevalences are aquagenic.The report of food source property also is seen in document.The infection rate of population of China Hepatitis E about 18%.Viral hepatitis type E accounts for 10% in the acute sporadic hepatitis, is one of China's Class B Notifiable disease.There is the time that continues by anti--HEVIgG after understanding hepatitis E virus (HEV) infection, resists-meaning of HEVIgG in Hepatitis E (viral hepatitis type E) is diagnosed with accurate, objective evaluation.
The diagnosis of viral hepatitis type E at present mainly is to detect anti-HEV antibody by ELISA (euzymelinked immunosorbent assay (ELISA)), and next has radiommunoassay (RIA), Western blotting to survey method, RT-PCR method, real-time fluorescence RT-PCR method etc.Western blotting is surveyed method sensitivity not as ELISA, RIA, RT-PCR and real-time fluorescence RT-PCR method, and the ELISA reagent sensitivity infects the window phase patient to HEV and can not diagnose ahead of time not as RIA, RT-PCR and real-time fluorescence RT-PCR reagent; Though and conventional RIA sensitivity is high, the half life period is short, forms radiocontamination easily; RT-PCR and real-time fluorescence RT-PCR method complex operation, poor stability, instrument costliness.Given this present invention has set up a kind of chemiluminescence enzyme promoting immunity analytical approach quick, responsive, high specificity and has detected hepatitis E virus antibody.The high sensitivity that methods such as radiommunoassay, fluoroimmunoassay are promptly arranged has overcome the shortcoming of these methods again.
Now, chemical luminescence immune analysis reagent box mostly is the luminous measuring system of the closed full-automatic chemical of import, apparatus expensive, and the use cost height is promoted the use of thereby limited, and can't be widely used in clinical diagnosis and research work.Kit of the present invention adopts the microwell plate chemiluminescence immunoassay technology, both can be applicable to the open luminous measuring instrument of semi-automatic chemistry, also can be used for full automatic measuring system, can realize fast detection the in enormous quantities, use cost is low, more can be adapted at large, medium and small hospital and apply.
Summary of the invention
The present invention adopts the indirect method principle, utilize the chemiluminescence enzyme immunoassay technology, with determinand generation immune response in enzyme-labelled antigen or antibody and the sample, enzyme on the formed immune complex remakes and is used for luminous substrate, the light signal strength that produces is directly proportional with the enzyme molecular number (activity) that participates in reaction, can judge the concentration of determinand or the variation of content according to luminous signal.
The objective of the invention is: the kit that a kind of chemiluminscence immunoassay hepatitis E virus IgG antibody is provided.Kit of the present invention comprises: hepatitis E virus IgG antibody positive and negative reference substance; Bag is by solid phase carrier; The enzyme labeling bond; Sample diluting liquid; Chemical luminous substrate liquid; And concentrated cleaning solution.
According to kit of the present invention, wherein, described solid phase carrier is microwell plate, plastic bead, plastic tube or magnetic-particle; Described marker enzyme is horseradish peroxidase or alkaline phosphatase; Described chemical luminous substrate is 1,2-two oxidative ethane analog derivatives, luminol or different luminol, wherein said 1,2-two oxidative ethane analog derivatives, be (diamantane)-1,2-two oxidative ethanes, 3-(2 '-the spiral diamantane)-4-methoxyl-4-(3 " the phosphorus acyloxy) phenyl-1,2-two oxidative ethanes (AMPPD), CSPD or CDP-Star.
An order more of the present invention provides the preparation method of kit of the present invention.Adopt the indirect method principle, utilize the chemiluminescence enzyme immunoassay technology, indirect method detects the hepatitis E virus IgG antibody in serum (slurry) sample.The preparation method of this invention kit is: with the reference substance of hepatitis E virus IgG antibody positive and negative serum preparation; With the antigen coated solid phase carrier of recombined hepatitis E hepatitis virus; With the anti-human IgG antibody of enzyme labeling (monoclonal antibody or how anti-); The preparation sample diluting liquid; Prepare the chemical luminous substrate liquid that above-mentioned enzyme acts on; The preparation concentrated cleaning solution; The above-mentioned each component of packing; And be assembled into finished product.
The method according to this invention, the step 2 of described solid phase carrier bag quilt) may further comprise the steps: be 9.0 dipotassium hydrogen phosphate (K with the 0.1MpH value
2HPO43H
2O) damping fluid is mixed with the antigen coated liquid of recombined hepatitis E hepatitis virus of desired concn, and coating buffer is carried on the solid phase carrier; Again with containing 0.1% casein, 0.01% gelatin, 0.1%Proclin300, the pH value is 7.0～7.5, concentration is that the phosphate buffer of 0.01M is as the above-mentioned solid phase carrier of confining liquid sealing.
In said method, and be microwell plate, plastic bead, plastic tube or magnetic-particle to described antigen coated solid phase carrier; Described bond is horseradish peroxidase, alkaline phosphatase; Described chemical luminous substrate is 1,2-two oxidative ethane analog derivatives, luminol or different luminol, wherein said 1,2-two oxidative ethane analog derivatives are (diamantane)-1,2-two oxidative ethanes, 3-(2 '-the spiral diamantane)-4-methoxyl-4-(3 " the phosphorus acyloxy) phenyl-1; 2-two oxidative ethanes, CSPD or CDP-Star, carried out preferred one by one.
Concrete, the mentioned reagent box comprises hepatitis E virus IgG antibody positive and negative reference substance, direct coated recombined hepatitis E hepatitis virus antigen microwell plate, sample diluting liquid, enzyme labeling thing, chemical luminous substrate liquid and concentrated cleaning solution etc.Wherein, described hepatitis E virus I gG antibody positive and negative reference substance is through HBsAg, anti-HIV, anti--TP and many parts of all negative pooled serums of anti-HCV TPPA, 60 ℃ of deactivations 1 hour, appropriateness dilution preparation (anti--HEVIgG antibody positive tire＞1: 1000); The microwell plate of bag quilt is the micropore lath in 48 or 96 holes, and the enzyme of labelled antibody is coupling horseradish peroxidase (HRP), and chemical luminous substrate liquid is luminol, and concentrated cleaning solution is PBST.
The present invention's " hepatitis E virus IgG antibody chemical luminescence immune assay determination reagent kit " can detect very single-mindedly and infect the hepatitis E virus IgG antibody in the human body after the hepatitis E virus, can judge result of treatment and change of illness state according to measuring hepatitis E virus IgG antibody situation of change.It has advantages such as easy, quick, sensitive, stable.This hepatitis E virus IgG antibody chemical luminescence is measured every index of kit all above the analysis level of ELISA kit, alternative RIA reagent and ELISA reagent.And detection system of the present invention is an open-sky technique, and is easy to be quick, do not need the expensive luminous measuring instrument of full-automatic chemical, is particularly suitable for vast middle and small hospital and promotes the use of.According to kit of the present invention, the antigen of bag quilt and the hepatitis E virus IgG antibodies of sample on the carrier, enzyme labeling two anti-again with solid phase on the antibodies that combines, so " indirect method " reaction pattern of the present invention's employing, what use is the enzymatic luminous substrate, by the chromogenic substrate in the light signal replacement EIA enzyme immunoassay that detects the luminous substrate generation, thereby having specificity, highly sensitive in EIA enzyme immunoassay reagent now, the diagnosis that can be hepatitis E virus IgG antibody provides more special, quick, reliable foundation.Kit of the present invention had both effectively utilized the chemiluminescence principle, had guaranteed the sensitivity that detects again.In addition, this pattern also is convenient to operation and is produced.
Embodiment
Embodiment 1 preparation hepatitis E virus IgG antibody chemical luminescence immune assay determination reagent kit of the present invention
In research process of the present invention, the present inventor has at first carried out screening experiment and Quality Identification to used starting material, comprises the luminous intensity of activity, chemical luminous substrate of the absorption property of activity, carrier (as lighttight white microwell plate) of labelled antibody and coated antibody and variation size, HRP and luminous duration etc.Then method for coating is studied, be cushioned liquid and protect liquid to experimentize, select optimal bag and be cushioned liquid and protection liquid, tested by concentration by the different bags of antibody and find best concentration conditions with different bags.Mark for HRP can have diverse ways, by explore repeatedly and the contrast experiment finally found easy, productive rate is high, cost is low, the reliable quality labeling method, and different enzyme dilutions has been carried out long-term investigation tested, made and can make the activity stabilized dilution of the long-term maintenance of enzyme labeling thing
One, enzyme labelled antibody preparation
Anti-human IgG antibody uses glutaraldehyde method and horseradish peroxidase, concrete steps comprise: get HRP10mg and be dissolved in 0.4ml, 0.25Mol/L in the pH6.8PBS liquid or in the 0.05Mol/L pH9.6 carbonate buffer solution, add 25% glutaraldehyde 0.1ml, add ice-cold analytically pure absolute ethyl alcohol 2ml behind 37 ℃ of incubation 2h, 2500r/min centrifugation 10min-15min, solution inclines.Precipitation is with 80% ethanol 4ml-5ml suspendible, and is the same centrifugal, and the ethanol that inclines will be managed inversion, and ethanol is fully flowed out.Precipitation adds the anti-IgG antibody-solutions of 0.5ml-1ml (containing about antibody 15mg) with the dissolving of 1ml 0.05Mol/L pH9.6 carbonate buffer solution, be placed on spend the night in the refrigerator after, add KH
2PO
4, the nearly neutrality of its pH value can be used, add equal-volume glycerine, preserve below-20 ℃.
It is selected that enzyme is marked anti-human IgG antibody's concentration
With the anti-human IgG antibody of enzymic-labelled antibody dilution with enzyme labeling, the dilution variable concentrations, adopting the square formation titrimetry to select the working concentration of enzyme labelled antibody is 1: 20000.
Enzyme mark monoclonal antibody dilution: based on 1000ml enzyme labelled antibody dilution, its component content is 2.9gNa2HPO412H2O, 0.3g NaH2PO42H2O, 9g NaCl, 25g BSA, 1ml Proclin300,1ml Tween-20, the weighing each component, in clean container, dissolve with distilled water, be adjusted to pH7.2～7.4, fixed molten 1000ml.
Two, the preparation of hepatitis E virus IgG antibody positive and negative reference substance
Mix more than 6 parts and detect hepatitis E virus I gG antibody positive serum or negative serum through the ELISA kit, through HBsAg, anti-HIV, anti--TP and anti-HCV negative antibody serum, 60 ℃ of deactivations 1 hour, filtration sterilization, packing, low temperature is frozen.
Three, the preparation of solid-phase coating plate
The bag quilt adds an amount of recombined hepatitis E hepatitis virus antigen mixing in coating buffer, add then in each hole of microwell plate, and every hole 100 μ L place absorption 24 hours for 4 ℃.
By solution, its each component content is 22.82g dipotassium hydrogen phosphate (K based on the 1000ml bag
2HPO43H
2O), in clean container, with distilled water dissolving, be adjusted to the fixed molten 1000ml in pH9.0～9.3.
Every hole adds confining liquid 120 μ L respectively, places 24 hours for 4 ℃.Get rid of confining liquid, on thieving paper, pat dry.Room temperature removal moisture drying 24 hours.Carry out envelope then.Place behind the envelope and checked not have gas leakage in 15 minutes, if there is gas leakage to need envelope again.2～8 ℃ of preservations of labeling postposition.
Based on the 1000ml lock solution, its each component content is 0.36gNaH
2PO
42H
2O, 2.76gNa
2HPO
412H
2O, 1g casein, 0.1g gelatin, 1mlProclin300, the weighing each component in clean container, with the distilled water dissolving, is adjusted to the fixed molten 1000ml of pH7.0.
Four, sample diluting liquid
Based on the 1000ml sample diluting liquid, its each component content is 0.59gNaH2PO42H2O, 5.8gNa2HPO412H2O, 30g NaCl, 10g BSA, 1mLTween-20,1mlProclin300, the weighing each component, in clean container, with the distilled water dissolving, transfer the fixed molten 1000ml in pH to 7.2～7.4.
Five, chemical luminous substrate A liquid
Based on 1000mlA liquid component content be: 10mM luminol 1.7716g, 0.3mM 4-xenol 0.051g, 0.05mM 4-iodobenzene boric acid 0.012g, boric acid 11.4g, borax 4.9g.The good each component amount of weighing is put into clean container, adds distilled water, the dissolving mixing, and adjusting the pH value is 8.0～10.0, constant volume 1000ml.
Six, chemical luminous substrate B liquid
Based on 1000mlA liquid component content be: 3.5mM urea peroxide 0.329g, 1ml 0.1%Tween20,51.58gNa
2HPO
412H
2O, 8.74g NaH
2PO
42H
2O, the good each component amount of weighing are put into clean container, add distilled water, the dissolving mixing, and adjusting the pH value is 7.0～7.6, constant volume 1000ml.
The good each component amount of weighing is put into clean container, adds distilled water, the dissolving mixing, and adjusting the pH value is 8.0～10.0, constant volume 1000ml.
A liquid mixes with B liquid equal proportion during use.
Seven, 20 times of cleansing solutions
Based on 20 times of cleansing solution each components of 1000ml content is 58g Na
2HPO
412H
2O, 2g NaH
2PO
42H
2O, 160g NaCl, 1mL Tween-20,1mL Proclin 300.The good each component amount of weighing is put into clean container, adds distilled water, the dissolving mixing, and adjusting the pH value is 7.2～7.4, constant volume 1000ml.
Eight, semi-manufacture and finished product are formed
The packing of above-mentioned steps products obtained therefrom is semi-manufacture.Semi-manufacture are by detecting: specificity, accuracy, sensitivity and stability are assembled into finished product behind the index assay approval.Could use after finished product kit measurement specificity after the assembling, accuracy, sensitivity and stable every index are qualified.
The present inventor has also carried out experimental study to the reaction pattern and the reaction conditions of kit, has finally determined the indirect method reaction pattern, and the influence of experimental result is tested with regard to the different reaction time, determines the optimal reaction time.By the influence experiment to measured value shows to the mensuration of luminous duration of chemical luminous substrate liquid and different fluorescent lifetime: be measured as the best between 5-25 minute after adding chemical luminous substrate liquid, its result is also comparatively accurate.
Embodiment 2～3 preparations hepatitis E virus IgG antibody chemical luminescence immune assay determination reagent kit of the present invention
Divided by the alkali phosphatase enzyme mark hepatitis E virus antibody, with AMPPD as chemical luminous substrate, with plastic bead, plastic tube as the pH value of carrier, coating buffer be 4.5, the pH value of confining liquid is outside 7.5, all the other all prepare hepatitis E virus IgG antibody chemical luminescence immune assay determination reagent kit with the method identical with embodiment 1.
Embodiment 4 preparations hepatitis E virus IgG antibody chemical luminescence immune assay determination reagent kit of the present invention
As solid phase carrier, adopt the EDC coupling method to prepare outside the magnetic particulate antigen solid phase carrier divided by magnetic-particle, all the other all prepare hepatitis E virus IgG antibody chemical luminescence immune assay determination reagent kit with the method identical with embodiment 1.
The using method of embodiment 5 kits of the present invention
The concrete operations of the hepatitis E virus IgG antibody chemical luminescence immune assay determination reagent kit of above embodiment 1 preparation are as follows:
1) in 4 ℃ of refrigerators, takes out kit, equilibrium at room temperature 15 minutes.
2) take out coated slab, insert on the grillage.
3) except that blank well, add each two hole of yin, yang reference substance in the reacting hole respectively, every hole 100ul, each hole adds sample 10ul to be checked, sample diluting liquid 100ul, with the micro-oscillator mixing that fully vibrates, 37 ℃ of incubations 0.5 hour.
4) get rid of dereaction liquid, wash plate 5 times, every hole adds enzyme-added then label 100 μ L, with the micro-oscillator mixing that fully vibrates, and 37 ℃ of incubations 0.5 hour.
5) get rid of dereaction liquid, wash plate 5 times, on clean thieving paper, buckle at last and do.
6) each hole adds each 50 μ L of chemical luminous substrate liquid A, B, and fully vibrating with micro-oscillator mixes, room temperature (20～25 ℃) lucifuge reaction 5 minutes.
7) must measure in the 5th～25 minute after adding chemical luminous substrate liquid, on the chemiluminescence measuring instrument, measure the luminous intensity (RLU) in each hole in regular turn, 1 second/hole of Measuring Time.
8) CO value=2.1N, RLU value 〉=CO value then sample is judged to be positive sample; RLU value＜CO value then sample is judged to be negative sample.
The methodology of embodiment 6 kits of the present invention is identified
According to manufacturing conventional in this area and vertification regulation the kit of preparation among the embodiment 1 is examined and determine, is seen the following form 1:
Sensitivity, specificity, accuracy and stability that " hepatitis E virus IgG antibody (anti--HEVIgG) chemical luminescence method immune analysis is measured kit " is described meet the calibrating requirement fully.
Embodiment 7 hepatitis E virus IgG antibody of the present invention (anti--HEVIgG) clinical practice and the ELISA kit of chemical luminescence method immune analysis mensuration kit compare
Clinical hospital use embodiment 1 hepatitis E virus IgG antibody (anti--HEVIgG) chemical luminescence method immune analysis is measured kit and the ELISA kit detects 652 parts of clinical samples, to contrast the coincidence rate that two kinds of kits detect.
One, the source of clinical serum specimen:
Hospital clinical is collected hepatitis E virus patient, non-hepatitis E virus patient and normal person's sample.
Two, use hepatitis E virus IgG antibody (anti--HEVIgG) chemical luminescence method immune analysis mensuration kit and the comparison of ELISA kit
1, method
Clinical hospital uses the embodiment of the invention 1 preparation " hepatitis E virus IgG antibody (anti--HEVIgG) chemical luminescence method immune analysis is measured kit " (by its instructions operation) and ELISA kit (by its instructions operation) to detect 652 parts of clinical samples (positive 105 examples of hepatitis B virus antigen, the HAAb positive 95, normal person's 245 examples, Hepatitis E IgG antibody positive 205 examples) respectively, to contrast the coincidence rate of two kinds of kits detections.
2, result
The clinical use embodiment 1 of hospital " hepatitis E virus IgG antibody (anti--HEVIgG) chemical luminescence method immune analysis is measured kit " and ELISA kit comparison and detection result (seeing Table 2)
Two kinds of methods of table 2. clinical hospital contrast relatively
Remarks:
Reagent of the present invention detects 1 routine Hepatitis E IgG antibody positive among the hepatitis A patient, through the clinical Hepatitis E IgG antibody positive of confirming as.Contrast agent detects the positive blood sample 1 routine Hepatitis E IgG antibody positive of hepatitis B, through the clinical feminine gender of confirming as.
The use of clinical hospital " hepatitis E virus IgG antibody (anti--HEVIgG) chemical luminescence method immune analysis is measured kit " and showing with ELISA kit comparison and detection result: the clinical detection index of kit of the present invention has has all met or exceeded the ELISA method, and kit specificity 100% of the present invention, susceptibility 100%.Kit of the present invention is detected hepatitis A patient 1 routine hepatitis E virus IgG antibody positive, through the clinical Hepatitis E IgG antibody true positives of confirming as; It is positive that contrast agent detects hepatitis B patient serum sample 1 example, through the clinical Hepatitis E IgG antibody false positive of confirming as.(anti--HEVIgG) chemical luminescence immune assay determination reagent kit specificity, susceptibility, anti-interference all are better than ELISA reagent to hepatitis E virus IgG antibody of the present invention, can be applied to clinical Hepatitis E medical diagnosis on disease.
Claims (9)
1, a kind of hepatitis E virus IgG antibody chemical luminescence immune assay determination reagent kit is characterized in that, described kit comprises: hepatitis E virus IgG antibody positive and negative reference substance; Bag is by solid phase carrier; The enzyme labeling bond; Sample diluting liquid; The chemical luminous substrate liquid that enzyme acted on; Concentrated cleaning solution.
2, kit as claimed in claim 1 is characterized in that, described bag is by microwell plate, plastic bead, plastic tube or the magnetic-particle of solid phase carrier for reorganization hepatitis E virus antigen direct coated.
3, kit as claimed in claim 1 is characterized in that, described enzyme labeling bond is the anti-human IgG (monoclonal antibody or how anti-) of horseradish peroxidase or alkali phosphatase enzyme mark.
4, kit as claimed in claim 1 is characterized in that, described chemical luminous substrate is 1,2-two oxidative ethane analog derivatives, luminol or different luminol.
5, kit as claimed in claim 1 is characterized in that, described chemical luminous substrate comprises A liquid and B liquid, wherein,
Described chemical luminous substrate A liquid comprises 10mM luminol, 0.3mM 4-xenol, 0.05mM 4-iodine substituted phenyl boric acid, 0.2M pH8.7 boric acid-borate buffer solution, and its pH value is 8.0～10.0;
Described chemical luminous substrate B liquid comprises 3.5mM urea peroxide, 0.1%Tween20,0.2M pH7.2 phosphate buffer, and its pH value is 7.0～7.6.
6, a kind of method for preparing the described kit of claim 1 is characterized in that may further comprise the steps: with the reference substance of hepatitis E virus IgG antibody positive and negative serum preparation; With recombined hepatitis E hepatitis virus antigen direct coated solid phase carrier; With the anti-human IgG antibody of enzyme labeling (monoclonal antibody or how anti-); The preparation sample diluting liquid; Prepare the chemical luminous substrate liquid that above-mentioned enzyme acts on; The preparation concentrated cleaning solution; The above-mentioned hepatitis E virus IgG antibody of packing positive and negative reference substance, sample diluting liquid, the anti-human IgG antibody of enzyme labeling, chemical luminous substrate liquid and the concentrated cleaning solution that this enzyme acted on; And be assembled into finished product.
7, method as claimed in claim 6 is characterized in that, described bag is by the step 2 of solid phase carrier) adopt following method: with 0.1M pH value 9.0 dipotassium hydrogen phosphate (K
2HPO43H
2O) damping fluid is mixed with the antigen coated liquid of recombined hepatitis E hepatitis virus of desired concn, and coating buffer is carried on the solid phase carrier; With containing 0.1% casein, 0.01% gelatin pH value is 7.0～7.5, and concentration is that the phosphate buffer of 0.01M seals above-mentioned solid phase carrier as confining liquid.
As claim 6 or 7 described methods, it is characterized in that 8, described bag is microwell plate, plastic bead, plastic tube or magnetic-particle by the solid phase carrier of hepatitis E virus antigen.
9, method as claimed in claim 6 is characterized in that, described enzyme is the anti-human IgG antibody (resisting or monoclonal antibody) of horseradish peroxidase or alkali phosphatase enzyme mark more; Described chemical luminous substrate is 1,2-two oxidative ethane analog derivatives, luminol or different luminol.Further, described 1,2-two oxidative ethane analog derivatives are (diamantane)-1,2-two oxidative ethanes, 3-(2 '-the spiral diamantane)-4-methoxyl-4-(3 " the phosphorus acyloxy) phenyl-1,2-two oxidative ethanes, CSPD or CDP-Star.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2008101032659A CN101551396A (en) | 2008-04-02 | 2008-04-02 | Chemiluminscence immunoassay kit of hepatitis E virus IgG antibody and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2008101032659A CN101551396A (en) | 2008-04-02 | 2008-04-02 | Chemiluminscence immunoassay kit of hepatitis E virus IgG antibody and preparation method thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN101551396A true CN101551396A (en) | 2009-10-07 |
Family
ID=41155752
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNA2008101032659A Pending CN101551396A (en) | 2008-04-02 | 2008-04-02 | Chemiluminscence immunoassay kit of hepatitis E virus IgG antibody and preparation method thereof |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102128928A (en) * | 2010-11-30 | 2011-07-20 | 中国人民解放军第二炮兵总医院 | Pepsase chemiluminescent immunoassay kit and preparation method thereof |
| CN102353777A (en) * | 2011-07-05 | 2012-02-15 | 深圳市卫武光明生物制品有限公司 | Kit and method for testing human cytomegalovirus IgG antibody |
| CN104090105A (en) * | 2014-07-10 | 2014-10-08 | 广州市丰华生物工程有限公司 | Method and kit for detecting hepatitis E virus (HEV) antibody and method for preparing kit |
| CN105445463A (en) * | 2014-09-25 | 2016-03-30 | 苏州新波生物技术有限公司 | Hepatitis E virus IgG antibody detection kit and preparation method and application thereof |
| CN105954510A (en) * | 2016-06-30 | 2016-09-21 | 深圳市亚辉龙生物科技股份有限公司 | Chemiluminescence immunodetection kit for anti-beta 2 glycoprotein I antibodies IgG and preparation method of kit |
-
2008
- 2008-04-02 CN CNA2008101032659A patent/CN101551396A/en active Pending
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102128928A (en) * | 2010-11-30 | 2011-07-20 | 中国人民解放军第二炮兵总医院 | Pepsase chemiluminescent immunoassay kit and preparation method thereof |
| CN102128928B (en) * | 2010-11-30 | 2013-10-09 | 中国人民解放军第二炮兵总医院 | Pepsase chemiluminescent immunoassay kit and preparation method thereof |
| CN102353777A (en) * | 2011-07-05 | 2012-02-15 | 深圳市卫武光明生物制品有限公司 | Kit and method for testing human cytomegalovirus IgG antibody |
| CN102353777B (en) * | 2011-07-05 | 2014-07-02 | 深圳市卫光生物制品股份有限公司 | Kit and method for testing human cytomegalovirus IgG antibody |
| CN104090105A (en) * | 2014-07-10 | 2014-10-08 | 广州市丰华生物工程有限公司 | Method and kit for detecting hepatitis E virus (HEV) antibody and method for preparing kit |
| CN105445463A (en) * | 2014-09-25 | 2016-03-30 | 苏州新波生物技术有限公司 | Hepatitis E virus IgG antibody detection kit and preparation method and application thereof |
| CN105954510A (en) * | 2016-06-30 | 2016-09-21 | 深圳市亚辉龙生物科技股份有限公司 | Chemiluminescence immunodetection kit for anti-beta 2 glycoprotein I antibodies IgG and preparation method of kit |
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