CN106755346A - A kind of nocardial kit of quick discriminating and its application method - Google Patents
A kind of nocardial kit of quick discriminating and its application method Download PDFInfo
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- CN106755346A CN106755346A CN201611090372.3A CN201611090372A CN106755346A CN 106755346 A CN106755346 A CN 106755346A CN 201611090372 A CN201611090372 A CN 201611090372A CN 106755346 A CN106755346 A CN 106755346A
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- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
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- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
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- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6844—Nucleic acid amplification reactions
- C12Q1/6851—Quantitative amplification
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Abstract
A kind of nocardial kit of quick discriminating, it is characterised in that：Probe implementation sequence is as follows：FAM CCTAGA TGCGCTTCTTGATGTCGAC TCTAGG DABCYL, kit forms are as follows：The design can be identified and be diagnosed without sequencing to Nocard's bacillus, solve the quick discriminating of pathogenicity actinomyces, the difficult neck bottle of identification and diagnosis, save appraisal cost and simple to operate, and do not interfere with the amplified reaction of PCR.
Description
Technical field
The present invention relates to a kind of nocardial kit of quick discriminating and its application method, belong to differential diagnosis kit
Research and development field.
Background technology
Nocardia（Nocardia）Be aerobic actinomyces, be saprophytic bacteria in soil, be not in humans and animals body just
Normal flora.The infection of promise card bacterium can both occur after Healthy People wound, and immune deficient patients' lethal infection can be caused again, work as suction
Or by wound enter human body after, the infection of lung and hypodermis can be caused, can also send out to whole body, cause various interior internal organs
Official infects.At least 25 kinds of the pathogen cured the disease to human or animal in Nocardia, separate from human body
Promise card bacterium, nocardia asteroides accounts for 90%, and Nocardia brasiliensis account for 7%, and nocardia caviae accounts for 3%.It has been reported that, star promise
Cattell bacterium is most common animal pathogen in aerobic actinomyces, and most common Animal diseases are bovine mastitis, are also once reported
Cause infection of marine fishes.Clinically only suspected there is corresponding infection potential, aerobic actinomyces would generally be failed to pinpoint a disease in diagnosis or
The detection method operations such as mistaken diagnosis, current conventional use of Nocard's bacillus is separately cultured, the round pcr of biochemical reactions and classics
It is cumbersome, time-consuming, sensitivity is low, is unsuitable for clinically rapid differential diagnosis.Therefore, nocardial quick, reliability, spy are set up
Different, sensitive and simple to operate detection method is very necessary.
The gene of gene diagnosis direct detection clinical samples pathogen infection, belongs to etiological diagnosis, in prevention from suffering from the diseases, diagnosis
And the everyway such as efficacy assessment is significant.The means such as classical use agarose gel electrophoresis are observed testing result
Analysis, human error is big, and false positive, false negative rate are high.Fluorescence polarization technology detects intensity of polarization light rather than fluorescence intensity, spirit
Sensitivity is high, quick, simple and accurate, is the technology for being used in particular for studying intermolecular interaction.This method is direct timely
Detect the combination/free rate of tracer molecule.Fluorescence polarization experiment is carried out in the solution supported without solid phase, it is allowed in as little as skin
The real balance of analysis in the range of mole level.Fluorescence polarization detects not doped samples, therefore sample can be processed again
And reanalysed and change influence to combining for evaluating pH, temperature and salinity.Additionally, fluorescence polarization experiment is real-time
Carry out, it is not limited to the research that balance is combined.TDI reactions are the prime end extensions under DNA template-directeds,
The identification for expanding, determining specific probe and target DNA by primer hybridize and mould pull instruct under Te Ding the special alkali bases of Wei Ge and mix
Enter it is triple check on, determine genotype, so that it is guaranteed that reaction specificity.
Specific gene be Nocard's bacillus exist most directly mark, have multiple studies have shown that, with nucleotide sequence
Based on classification of actinomycetes and differentiate research, 16S rRNA sequences are difficult to differentiate between some kinds, and house-keeping genesecA1
Sequence there is enough conservatives and changeability, can be used as distinguishing target molecule not of the same race.Sec System is quite conservative
Mechanism of secretion, all exist in ancient bacterium, bacterium and eucaryote.Sec Secretory pathway in bacterium, especially Escherichia coli
Extensive research is obtained.secA1Albumen（Tool ATPase activity）As the important component of precursor protein translocase, it is
Across the plasma membrane conveying of albumen provides driving force.Have recently multiple studies have shown that, in actinomyces based on nucleotide sequence point
In class and discriminating research, 16S rRNA sequences are difficult to differentiate between some and plant, and house-keeping genesecA1 Sequence there is enough guarantors
Keeping property and changeability, can be used as differentiation target molecule not of the same race.
Fischer etc. has expanded 29 the 47 of kind plants of ATCC standard bacterias of Mycobacterium and 59 plants are clinically separated bacteriumsecA1 Gene order, finds that secretion related gene can be used for distinguishing the different strain in Mycobacterium first.
Conville etc. analysis compare Nocardia 29 kinds 16S rRNA andsecA1Gene order, findssecA1
Gene can more preferably more accurately distinguish between and differentiate the strain in Nocardia.Similarly, we are in 23 belonged to Ge Dengshi
Type sepecies are carried out respectivelysecA1The comparative analysis of gene and 16S rRNA Phylogenetics, then draws a conclusion,secA1
It is used for the identification for carrying out kind of level with the research of phyletic evolution as house-keeping gene, is ideal target.
The content of the invention
The technical problem to be solved in the present invention：A kind of molecular beacon probe for being capable of specific amplification of design, using real glimmering
Fluorescent Quantitative PCR method, so that research and develop that nocardial diagnostic kit in the aerobic actinomyces of common clinical pathogenic can be differentiated, can
To overcome the difficult neck bottle of the quick discriminating, identification and diagnosis of present pathogenicity actinomyces.
The technical scheme is that：A kind of nocardial kit of quick discriminating, molecular beacon probe implementation sequence
It is as follows：FAM-CCTAGATGCGCTTCTTGATGTCGAC TCTAGG-D A BCYL, kit forms are as follows：
| 1 | PCR Mix | 1.0mL × 5 bottle |
| 2 | secA1Sense primer | 0.5 mL × 1 bottle |
| 3 | secA1Anti-sense primer | 0.5 mL × 1 bottle |
| 4 | Molecular beacon probe | 0.5 mL × 1 bottle |
| 5 | Distilled water | 1.0mL × 5 bottle |
| 6 | Aseptic eight comb | 8 × 7 |
| 7 | Aseptic eight combs lid | 8 × 7 |
| 8 | Specification | 1 part |
A kind of application method of the nocardial kit of quick discriminating, using 20 μ L systems：The μ L of PCR Mix 10, upstream and downstream is drawn
Each 1 μ L of thing, the μ L of DNA profiling 1, the μ L of molecular beacon probe 1, the μ L of distilled water 6；Real-time fluorescence quantitative PCR then is carried out, its amplification bar
Part is：Predegeneration：94℃ 4min；1. it is denatured：94 DEG C, 30s；2. anneal：58 DEG C, 50s；3. extend：72 DEG C, 90s； ①②
3. circulate 40 times, last 72 DEG C of extensions 10min.
According to the requirement that general cross probe is designed, according to the Nocardia announced on GenBankSecA1Gene sequence
Row, using Primer Software for Design molecular beacon probes, and the 19bp that will be chosen DNA fragmentation and Nocardia bacterial strain base
Because group DNA sequence dna carries out Homology search, it is determined that being one section of specific DNA without homologous fragment.Set according still further to molecular beacon probe
The requirement of meter, 6 alkali complementary and unrelated with target DNA are added at 5 ' and 3 ' ends of 19bp DNA fragment specifics respectively
Base arm, 5 ' ends are CCTAGA, and 3 ' ends are TCTAGG, form a hairpin structure（Such as Fig. 2）, 19 bp specific DNAs are constituted to be sent out
The Lu of folder flutters ring, and 6 complementary bases match the stem to form hair clip.In 5 ' end mark fluorescent element FAM of hairpin structure, 3 ' end marks
Note quencher DABCYL.
Beneficial effects of the present invention：
Domestic scholars Liu Lu etc. has invented a kind of Yellowtail fishes Nocard's bacillus fluorescent quantificationally PCR detecting kit and detection method, the inspection
Test agent box can detect the Yellowtail fish Nocard's bacillus of early stage with sensitive, quick, quantitative, specific, but cannot be used for clinical disease
People infects nocardial quick detection；And clinician is usually used in detecting that the kit of the aerobic actinomyces of cause of disease can be 6 at present
The clinical 17 kinds of common mycobacterias of detection in hour：Mycobacterium tuberculosis complex, Mycobacterium intracellulare, mycobacterium avium,
Mycobacterium gordonae, mycobacterium kansasii, mycobacterium fortuitum, Mycobacterium scrofulaceum, pale yellow mycobacteria, native branch bar
Bacterium, tortoise-mycobacterium abscessus, Mycobacterium graminis, achromatic mycobacterium, sea-mycobacterium buruli, gold mycobacteria, Soviet Union
Er Jia-Ma Ermo mycobacterias, mycobacterium xenopi and mycobacterium smegmatis, but Nocard's bacillus can not be detected.
Compared with the prior art, the present invention is by the detectable Nocard's bacillus of designsecA1Gene molecule beacon probe, can not
It can be identified and is diagnosed with sequencing, be solved the difficult neck of the quick discriminating of pathogenicity actinomyces, identification and diagnosis
Bottle, saves appraisal cost and simple to operate.
Inventor find, for Actinomycetal in several main related Pseudomonas such as Nocardia, Gordona, divide
Ramibacterium, streptomyces etc.SecA1Gene translation amino acid sequence out, not belonging to other strain together respectively has its specificity
Motif（motif）.According to the discovery in our early-stage Studies, can be designed that specificity amplification primer and probe, using point
Its specific primer is applied in the method for sub- beacon probe combination real-time fluorescence quantitative PCR, research and development in different reaction systems,
The diagnostic kit of the strain of synchronous detection and identification difference actinomyces so that, reliability quick to the clinical detection of pathogen, spy
It is different and simple to operate, the rate of missed diagnosis and testing cost of multiple mixed infection can be especially reduced, checkability can be improved, with sequencing
Detection method coincidence rate is high, and susceptibility is high.
Brief description of the drawings
Fig. 1 is the real-time fluorescence PCR result of different strains；A：Nocard's bacillus CMCC (F) D4C secA1Gene magnification knot
Really；B：Gordonia bronchialis IFM10866 secA1Gene magnification result；C：Rhodococcus equi CMCC (F) D4B secA1Gene magnification knot
Really；D：Negative control（Nocard's bacillus CMCC (F) D4C is not added with probesecA1Gene magnification result）；E：Blank（Water adds spy
Pin）；
Fig. 2 is molecular beacon probe schematic diagram；
Fig. 3 is Nocard's bacillussecA1Gene PCR amplification；M：DL2000；1-5：Nocard's bacillus CMCC (F) D4CsecA1Base
Gene-amplification result；
Fig. 4 is the influence that molecular beacon probe is expanded to PCR；M：DL2000；1：Nocard's bacillus CMCC (F) D4CsecA1Gene
Amplification；2：1 is repeated to test；3：After Nocard's bacillus CMCC (F) D4C+ molecular beacon probessecA1Gene magnification result； 4：
3 are repeated to test.
Specific embodiment
1. materials and methods
1.1 experimental strains and reagent
Nocardia bacterial strain CMCC (F) D4C, Rhod bacterial strain CMCC (F) D4B and Gordona bacterial strain IFM10866 are each
One plant, bacterial strain is given by dept. of dermatology of Shanghai Long March Hospital and Institute of Microorganism, Academia Sinica.
1.2 instruments
ABI StepOnePlus real-time fluorescence quantitative PCRs instrument is the silent winged generation that science and technology of match（China）Co., Ltd's product.
The design of 1.3 molecular beacon probes and synthesis
According to the requirement that general cross probe is designed, according to the Nocardia announced on GenBanksecA1Gene order,
Using Primer Software for Design molecular beacon probes, and the 19bp that will be chosen DNA fragmentation and Nocardia strain gene group
DNA sequence dna carries out Homology search, it is determined that being one section of specific DNA without homologous fragment.According still further to molecular beacon probe design
It is required that, 6 base arms complementary and unrelated with target DNA are added respectively at 5 ' and 3 ' ends of 19bp DNA fragment specifics,
5 ' ends are CCTAGA, and 3 ' ends are TCTAGG, form a hairpin structure（Such as Fig. 2）, the Lu of 19 bp specific DNAs composition hair clip
Ring is flutterred, 6 complementary bases match the stem to form hair clip.In 5 ' end mark fluorescent element FAM of hairpin structure, 3 ' end marks are quenched
Agent DABCYL.
Molecular beacon probe is synthesized by Li Fei Bioisystech Co., Ltd, and the probe of synthesis is purified with HPLC：A liquid
（0.1mol/L acetic acid triacetyls）80%-30%, B liquid（75% acetonitrile+A liquid）20%-70%, gradient elution 25min, then use B liquid
（100%）5min, flow velocity 1mL/min are washed, corresponding peak is collected, purity molecular beacon probe higher is obtained.
It is prepared by 1.4 templates
The template of negative control and positive control is processed as follows：Each bacterium is in after 37 DEG C of incubator cultures 5-7 days：1. picking
0.05g wet thallus, mix after adding 35 μ L lysates and 15 μ L 20%SDS solution；2. microwave 90s;3. 459 μ L are rapidly joined to take out
Extract, mixes；4. 500 μ L phenol chloroform liquid are added, is mixed, 13400g is centrifuged 10min, as far as possible Aspirate supernatant；5. repeat
Step is 4. once；6. take supernatant and add 800 μ L absolute ethyl alcohols and 80 μ L 3M sodium acetates（pH4.8-5.2）Afterwards, it is stored at room temperature
10min;7. supernatant is removed after 13400g centrifugations 10min, with 70% ethanol（150-200μL）Wash 1-2 times；8. supernatant is removed in centrifugation, in
37-55 DEG C of incubator drying；9. plus 1 × TE of 30-50 μ L dissolving after be put into -20 DEG C it is standby.
The amplification of 1.5 real-time fluorescence quantitative PCRs
Using 20 μ L systems：The μ L of PCR Mix 10, each 1 μ L of upstream and downstream primer, the μ L of DNA profiling 1, the μ L of molecular beacon probe 1, double steamings
The μ L of water 6, if blank.Routinely PCR conditions are expanded：After 95 DEG C of 5min, 95 DEG C of 1min, 63 DEG C of 1min, 72 DEG C of 1min,
35 circulations, 72 DEG C of 10min.It is repeated 3 times amplification experiment.
2. result
The availability of 2.1 molecular beacon probes
The molecular beacon probe of this research and design is used to detect Nocard's bacillussecA1Gene, sends out after being detected through real-time fluorescence PCR
It is existing：Only Nocard's bacillus produces positive signal, and remaining aerobic actinomyces is negative signal（See Fig. 1）.Illustrate the use of this research and design
In detection Nocard's bacillussecA1The molecular beacon probe of gene can it is quick, special, delicately detect Nocard's bacillus, can be with other
Non- Nocard's bacillus differentiates to be distinguished.
The influence that 2.2 molecular beacon probes are expanded to PCR
Template is expanded as PCR using the bacterial strain containing Nocard's bacillus strain specific DNA respectively, two experiments are designed, one is in PCR
Molecular beacon probe is added in reaction solution, another experiment is free of probe in PCR reaction solutions, while entering performing PCR amplification.Agarose
Gel electrophoresis results show（Fig. 4）, probe and the amplified band without probe are added in PCR reaction solutions generally within same position
Put, the amount of amplified production is also basically identical, this shows to add molecular beacon probe not interfere with the expansion of PCR in PCR amplification liquid
Increase reaction.
Claims (3)
1. the nocardial kit of a kind of quick discriminating, it is characterised in that：Molecular beacon probe implementation sequence is as follows：FAM-
CCTAGATGCGCTTCTTGATGTCGAC TCTAGG-DABCYL, kit forms are as follows：
。
2. the nocardial kit of a kind of quick discriminating according to claim 1, it is characterised in that：Described molecule letter
Mark the design of probe：According to the Nocardia announced on GenBanksecA1Gene order, using Primer Software for Design
Molecular beacon probe, and the DNA fragmentation of the 19bp of selection and Nocardia strain gene group DNA sequence are carried out into homologous ratio
Compared with determining one section of specific DNA without homologous fragment；According still further to the requirement of molecular beacon probe design, in 19bp specificity
Complementary and 6 base arms unrelated with target DNA are added at the 5 ' of DNA fragmentation and 3 ' ends respectively, and 5 ' ends are CCTAGA, and 3 ' ends are
TCTAGG, forms a hairpin structure, and the Lu that 19bp specific DNAs constitute hair clip flutters ring, and 6 complementary bases match to form hair clip
Stem, in 5 ' end mark fluorescent element FAM of hairpin structure, 3 ' ends mark quencher DABCYL.
3. a kind of application method of the nocardial kit of quick discriminating as claimed in claim 1 or 2, it is characterised in that：
Using 20 μ L systems：The μ L of PCR Mix 10, each 1 μ L of upstream and downstream primer, the μ L of DNA profiling 1, molecular beacon probe 1 μ L, the μ of distilled water 6
L；Real-time fluorescence quantitative PCR then is carried out, its amplification condition is：Predegeneration：94℃ 4min；1. it is denatured：94 DEG C, 30s；2. move back
Fire：58 DEG C, 50s；3. extend：72 DEG C, 90s；1. 2. 3. circulate 40 times, last 72 DEG C of extensions 10min.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112609011A (en) * | 2020-12-09 | 2021-04-06 | 北京华瑞康源生物科技发展有限公司 | Nocardia nucleic acid fluorescence real-time PCR detection primer, probe, kit and method |
| CN113186319A (en) * | 2021-06-22 | 2021-07-30 | 广东海洋大学 | Primer, kit and method for detecting Nocardia seriolae |
| CN115746112A (en) * | 2022-07-14 | 2023-03-07 | 山东第一医科大学附属省立医院(山东省立医院) | Nocardia specific antigen protein, serological diagnosis kit and application thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1904064A (en) * | 2006-06-15 | 2007-01-31 | 南开大学 | Star shaped nocardia multiple PCR fast detection kit and detection method |
| CN101962678A (en) * | 2010-09-26 | 2011-02-02 | 宁波大学 | Nocardia seriolae fluorescence quantitative PCR detection kit and detection method |
| US20140094383A1 (en) * | 2012-10-02 | 2014-04-03 | Ohio State Innovation Foundation | Tethered Lipoplex nanoparticle Biochips And Methods Of Use |
| CN104263844A (en) * | 2014-10-21 | 2015-01-07 | 中国疾病预防控制中心传染病预防控制所 | Nucleotide sequence for detecting Nocardia calcarea and application thereof |
-
2016
- 2016-12-01 CN CN201611090372.3A patent/CN106755346A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1904064A (en) * | 2006-06-15 | 2007-01-31 | 南开大学 | Star shaped nocardia multiple PCR fast detection kit and detection method |
| CN101962678A (en) * | 2010-09-26 | 2011-02-02 | 宁波大学 | Nocardia seriolae fluorescence quantitative PCR detection kit and detection method |
| US20140094383A1 (en) * | 2012-10-02 | 2014-04-03 | Ohio State Innovation Foundation | Tethered Lipoplex nanoparticle Biochips And Methods Of Use |
| CN104263844A (en) * | 2014-10-21 | 2015-01-07 | 中国疾病预防控制中心传染病预防控制所 | Nucleotide sequence for detecting Nocardia calcarea and application thereof |
Non-Patent Citations (3)
| Title |
|---|
| FANRONG KONG等: "secA1 Gene Sequence Polymorphisms for Species Identification of Nocardia Species and Recognition of Intraspecies Genetic Diversity", 《JOURNAL OF CLINICAL MICROBIOLOGY》 * |
| PATRICIA S. CONVILLE等: "Analysis of secA1 Gene Sequences for Identification of Nocardia Species", 《JOURNAL OF CLINICAL MICROBIOLOGY》 * |
| 孙渭歌: "诺卡菌多重PCR及Real-time PCR检测方法的建立", 《中国优秀硕士学位论文全文数据库 医药卫生科技辑》 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112609011A (en) * | 2020-12-09 | 2021-04-06 | 北京华瑞康源生物科技发展有限公司 | Nocardia nucleic acid fluorescence real-time PCR detection primer, probe, kit and method |
| CN113186319A (en) * | 2021-06-22 | 2021-07-30 | 广东海洋大学 | Primer, kit and method for detecting Nocardia seriolae |
| CN113186319B (en) * | 2021-06-22 | 2022-06-28 | 广东海洋大学 | Primer, kit and method for detecting Nocardia seriolae |
| CN115746112A (en) * | 2022-07-14 | 2023-03-07 | 山东第一医科大学附属省立医院(山东省立医院) | Nocardia specific antigen protein, serological diagnosis kit and application thereof |
| CN115746112B (en) * | 2022-07-14 | 2023-10-13 | 山东第一医科大学附属省立医院(山东省立医院) | Nocardia specific antigen protein, serological diagnosis kit and application thereof |
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