CN102399903B - Chikungunya virus isothermal amplification detection kit and primer thereof - Google Patents
Chikungunya virus isothermal amplification detection kit and primer thereof Download PDFInfo
- Publication number
- CN102399903B CN102399903B CN2011103019932A CN201110301993A CN102399903B CN 102399903 B CN102399903 B CN 102399903B CN 2011103019932 A CN2011103019932 A CN 2011103019932A CN 201110301993 A CN201110301993 A CN 201110301993A CN 102399903 B CN102399903 B CN 102399903B
- Authority
- CN
- China
- Prior art keywords
- primer
- seq
- chikungunya virus
- reaction
- detection
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Images
Classifications
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Abstract
The invention provides a chikungunya virus isothermal amplification detection kit and a primer thereof. The kit comprises the primer, Bst DNA polymerase, a reaction liquid and SYBR Green I fluorescent dye, wherein the reaction liquid is 10mM deoxyribonucleoside triphosphate, 10*reaction buffer liquid and 150mM MaSO4; the sequences of the primer are sequences represented by SEQ ID NO:1-4 or sequences represented by SEQ ID NO:5-8 or sequences represented by SEQ ID NO:9-12 or sequences represented by SEQ ID NO:13-16. The kit provided by the invention has the advantages of short detection time, strong specificity, high sensitivity, high detection accuracy and programmed reaction, is easy and convenient to qualify and is widely applied to conventional detection and epidemiological survey in clinic and ports.
Description
Technical field
The present invention relates to the molecular Biological Detection field, be specifically related to a kind of Chikungunya virus isothermal amplification fast detecting reagent kit and primer thereof.
Background technology
Chikungunya virus is the pathogenic agent that causes the chikungunya pyreticosis, mainly bites propagation by yellow-fever mosquito.Chikungunya heat is a kind of Amphixenosis, this disease be generate heat, fash and arthralgia be the viral acute infectious disease of principal character, mainly be popular in Africa and south east asia, though lethality rate is very low, the area higher in mosquito matchmaker density forms large-scale outbreak and popular easily.Yellow-fever mosquito extensively distributes on China south China, southwest and other places, becomes the potential inducement of the outbreak of epidemic of this disease.Only the hot suspected case of chikungunya of India's report in 2006 surpasses 1,390,000, and the sickness rate of some areas surpasses 45%.Detected China's the first Introduced cases Chikungunya virus case by Guangdong Entry-Exit Inspection and Quarantine Bureau in 2008, this prompting is along with the population mobility strengthens, and Chikungunya virus is in the popular possibility that become of China.Therefore, a cover is quick, sensitive, high specificity, and easy and simple to handle, does not rely on the detection method of valuable instrument and equipment, can submit necessary information for prevention and control, the epidemiology survey of this disease.
At present Chikungunya virus being detected has several different methods, comprises that viral separation and Culture is gene testers such as the immunology detection technology, nucleic acid probe, polymerase chain reaction (PCR) technology of main microbiology diagnostic method, specific antibody.Wherein the cause of disease detection of nucleic acids all improves a lot at aspects such as rapidity, security, accuracy and susceptibilitys, these new technologies do not need microorganism is separated purification, and directly with sample or sample nucleic acid extractive its gene and gene product are carried out rapid detection, and combine with Protocols in Molecular Biology and information biology means, to accurately, fast, the direction of sensitivity and automatization develops.
PCR (polymerase chain reaction) method is most popular nucleic acid amplification method so far, and it brings into play big effect emphatically as a kind of gene amplification technology of simple and effective in the pathogenic bacteria diagnostic procedure.Although PCR method operates relatively simple, amplified production can be assembled at short notice in a large number.Except PCR method, also have other nucleic acid constant-temperature amplification methods, such as amplification of nucleic acid sequences method (NASBA), from sequence transfer printing (3SR) and strand displacement transfer printing (SDA) etc.The detection level of these three kinds of methods all is no less than 10 copies, in about the 1 hour consuming time similar scope that just target nucleic acid can be increased.The immunology detection technology is fast and convenient with low cost, also can be as the detection means of Chikungunya virus.
Principle according to round pcr, sex change, annealing, three steps of extension need be arranged, the temperature of reaction of each step and time are all inequality and very accurate requirement arranged, even need carry out 20-40 circulation and finish, therefore need to rely on valuable relatively PCR temperature cycler and control, be unfavorable for on-site diagnosis requirement.Amplification of nucleic acid sequences method (NASBA), from sequence transfer printing (3SR) and strand displacement transfer printing (SDA) though belong to the isothermal duplication method, they are not strong to the specific amplification of target sequence, also need follow-up experimental implementation means to detect for amplified production after the feasible amplification, therefore have the shortcoming of complex operation.The monoclonal antibody of immunology detection technical requirements high quality high stability, otherwise accuracy is not enough; In addition, because the Chikungunya virus genome is shorter, only be 11.8kb, so expressed proteins and other correlated virus, for example o'nyong-nyong virus produces cross reaction, so specificity is relatively low.
In a word, at present, the detection technique of Chikungunya virus mainly contains: viral separation and Culture, patients serum's antibody test and RT-PCR gene test.Virus is separated the gold standard that belongs to present detection, but because there is the long problem of sense cycle in this technology, is unfavorable for the requirement of rapid detection; Serum Antibody Detection can the quick diagnosis Chikungunya virus, but produces cross reaction because this antiviral antibody is easy to o'nyong-nyong virus, thereby specificity is relatively low; Though the RT-PCR technology can be good at addressing the above problem, owing to need to rely on valuable relatively plant and instrument, be unfavorable for the requirement that field diagnostic and basic unit are detected.
Dna circle mediated constant temperature nucleic acid amplification technology (Loop-mediated Isothermal Amplification of DNA, abbreviation LAMP) overcomes the deficiency of gene amplification method in the past, can be special under constant temperature, carry out nucleic acid amplification efficiently, rapidly, have a lot of superiority.This technology has the following advantages: (1) high specificity, and the PCR reaction only needs a pair of primer to identify target DNA, reaches the purpose of Molecular Detection, and the LAMP technology is used 2 pairs of primers, identify 6 different zones of target DNA altogether, so specificity is higher; (2) highly sensitive, because this technology has been used a kind of sensitivity, archaeal dna polymerase that amplification efficiency is high, therefore improved sensitivity greatly, and shortened detection time; (3) simple to operate, be with the naked eye just can judge the result, and under isothermal condition, react, the valuable instrument and equipment that do not rely on easy and simple to handle.
Summary of the invention
The objective of the invention is the defective at above-mentioned prior art, a kind of Chikungunya virus isothermal amplification detection kit and primer thereof are provided, reach weak point detection time, high specificity, highly sensitive, effect that pertinency factor is high, make its response proceduresization, be widely used in conventional sense and epidemiology survey clinical, the port.
To achieve these goals, technical scheme of the present invention is as follows:
A kind of Chikungunya virus isothermal amplification detection kit of the present invention, described test kit comprise primer, Bst archaeal dna polymerase, reaction solution and SYBR Green I fluorescence dye; Described reaction solution is 10mM deoxynucleoside triphosphate, 10 * reaction buffer and 150mM MgSO4; Described primer sequence is the sequence shown in SEQ ID NO:1-4:
Outer primer 1:CGCCCTCTTTAACGGACATG;
Outer primer 2:AATTCGGCGCTGGCTAAG;
Inner primer 1:TGCCTTTCTTGCTGGCTGCATATACCAGCCTGCACCCATT;
Inner primer 2:AGTGTGCGGTGCATTCGATGATGCAGCTGAGAATTCCCTTC;
Or the sequence shown in SEQ ID NO:5-8:
Outer primer 3:GCTGAAAACACGCAGTTGAG;
Outer primer 4:TGGCCCCACAATGAATTTGG;
Inner primer 3:GCGGTATGAGCCCTGTATGCTGAAGCACATGTGGAGAAGTCC;
Inner primer 4:CAGCTAAGCTCCGCGTCCTTTCGCCGTTTGCATAGGC;
Or the sequence shown in SEQ ID NO:9-12:
Outer primer 5:CAGGCACCATCTGGCTTTA;
Outer primer 6:CCCCCAAAGTCTGAGGAATG;
Inner primer 5:GTTCCCTACGGCGCAGTTCAGCAGCACACAGCACCATT;
Inner primer 6:GCCCATCTCCATCGACATACCGCGCACGACATGTCCGTTAA;
Or the sequence shown in SEQ ID NO:13-16:
Outer primer 7:CGAAGCACATGTGGAGAAGT;
Outer primer 8:AGACGTCACCTTTGTACACC;
Inner primer 7:TTGGTAAAGGACGCGGAGCTTTTGCATCAGCATACAGGGC;
Inner primer 8:ACTGCCTATGCAAACGGCGAGTCCAGGCTGAAGACATTGG.
Use the detection kit of the invention described above that Chikungunya virus is detected, method is as follows:
(1) to the pre-treatment of test sample: with conventional method rapid extraction sample rna;
(2) according to following formulated reaction solution: reaction system is inner primer 1 and the inner primer 2 of 25uL:1-10mM, 0.1-10mM outer primer 1 and outer primer 2 (wherein inner primer 1, inner primer 2, outer primer 1 and outer primer 2 are the covers in the quadruplet primer), the dNTPs of 1-10mM, the MgSO4 of 1-10mM, the trimethyl-glycine of 1-10M, 1-5uL are slightly carried DNA, 1-8U Bst archaeal dna polymerase, 1-8U ThermoScript II, adding distil water be to 25uL, gentle mixing.
(3) carry out the loop-mediated isothermal amplification reaction: carried out the endless chain replacement(metathesis)reaction in 0.5 to 1.5 hour 60～65 ℃ of insulations.
(4) analysis and judgement reaction product result: add the fluorescence dye (SYBR Green I) that sell fluorescence dye such as market in reaction product, mixing leaves standstill 5min, and the reaction product shows green is then positive, shows orange then negative.
The present invention also provides a kind of primer for the detection of Chikungunya virus isothermal duplication, and this primer is designed at the special E1 gene of Chikungunya virus, and described primer sequence is the sequence shown in SEQ ID NO:1-4:
Outer primer 1:CGCCCTCTTTAACGGACATG;
Outer primer 2:AATTCGGCGCTGGCTAAG;
Inner primer 1:TGCCTTTCTTGCTGGCTGCATATACCAGCCTGCACCCATT;
Inner primer 2:AGTGTGCGGTGCATTCGATGATGCAGCTGAGAATTCCCTTC;
Or the sequence shown in SEQ ID NO:5-8:
Outer primer 3:GCTGAAAACACGCAGTTGAG;
Outer primer 4:TGGCCCCACAATGAATTTGG;
Inner primer 3:GCGGTATGAGCCCTGTATGCTGAAGCACATGTGGAGAAGTCC;
Inner primer 4:CAGCTAAGCTCCGCGTCCTTTCGCCGTTTGCATAGGC;
Or the sequence shown in SEQ ID NO:9-12:
Outer primer 5:CAGGCACCATCTGGCTTTA;
Outer primer 6:CCCCCAAAGTCTGAGGAATG
Inner primer 5:GTTCCCTACGGCGCAGTTCAGCAGCACACAGCACCATT;
Inner primer 6:GCCCATCTCCATCGACATACCGCGCACGACATGTCCGTTAA;
Or the sequence shown in SEQ ID NO:13-16:
Outer primer 7:CGAAGCACATGTGGAGAAGT;
Outer primer 8:AGACGTCACCTTTGTACACC;
Inner primer 7:TTGGTAAAGGACGCGGAGCTTTTGCATCAGCATACAGGGC;
Inner primer 8:ACTGCCTATGCAAACGGCGAGTCCAGGCTGAAGACATTGG.
The present invention uses Bioinformatics Platform to carry out extensive genome analysis; introduce degeneracy base and genetic component type processing for pleomorphism site; make design of primers more perfect; when design of primers; singularity according to the LAMP technology; take into full account primer dimer Δ G, 3 ' and 5 ' terminal Δ G, mispairing probability, optimized primer spacing and amplification efficiency, improved specificity and the sensitivity of amplified reaction greatly.
The invention has the beneficial effects as follows: compared with prior art, 1. only need just energy amplified reaction of a steady temperature, do not need special reagent and equipment; And, within a reaction system, can carry out the amplification of reverse transcription and target gene, handled easily simultaneously; 2. high specific: use six sections, four primers, whether the existence that just can judge target substance according to whether increasing to be, and false positive rate is 0; 3. fast efficient amplification: amplification only needed to finish in 1 hour, and the productive rate height; 4. highly sensitive: use the lowest detection limit to Chikungunya virus to reach in 10 copies, the recall rate of sample reaches high to 99%; 5. identify easy: identify by visual inspection, need not other any analytical procedures such as electrophoresis.6. purposes is wide, can be widely used in conventional sense and epidemiology survey clinical, the port.
Description of drawings
Fig. 1 is the colour-change figure as a result that detects the reaction tubes of Chikungunya virus with test kit of the present invention.
Embodiment
The invention will be further described below in conjunction with the drawings and specific embodiments, but not as a limitation of the invention.
Embodiment 1: Chikungunya virus isothermal gene amplification fast detecting kit
Test kit of the present invention is composed of the following components:
(1) detects primer
(2) archaeal dna polymerase: Bst archaeal dna polymerase
(3) reaction solution: 10mM dNTP (deoxynucleoside triphosphate), 10 * ThermoPol Buffer (reaction buffer), 150mMMgSO4 (sal epsom)
(4) fluorescence dye: SYBR Green I
The detection primer sequence is the sequence shown in SEQ ID NO:1-4:
Outer primer 1:CGCCCTCTTTAACGGACATG;
Outer primer 2:AATTCGGCGCTGGCTAAG;
Inner primer 1:TGCCTTTCTTGCTGGCTGCATATACCAGCCTGCACCCATT;
Inner primer 2:AGTGTGCGGTGCATTCGATGATGCAGCTGAGAATTCCCTTC;
Or the sequence shown in SEQ ID NO:5-8:
Outer primer 3:GCTGAAAACACGCAGTTGAG;
Outer primer 4:TGGCCCCACAATGAATTTGG;
Inner primer 3:GCGGTATGAGCCCTGTATGCTGAAGCACATGTGGAGAAGTCC;
Inner primer 4:CAGCTAAGCTCCGCGTCCTTTCGCCGTTTGCATAGGC;
Or the sequence shown in SEQ ID NO:9-12:
Outer primer 5:CAGGCACCATCTGGCTTTA;
Outer primer 6:CCCCCAAAGTCTGAGGAATG;
Inner primer 5:GTTCCCTACGGCGCAGTTCAGCAGCACACAGCACCATT;
Inner primer 6:GCCCATCTCCATCGACATACCGCGCACGACATGTCCGTTAA;
Or the sequence shown in SEQ ID NO:13-16:
Outer primer 7:CGAAGCACATGTGGAGAAGT;
Outer primer 8:AGACGTCACCTTTGTACACC;
Inner primer 7:TTGGTAAAGGACGCGGAGCTTTTGCATCAGCATACAGGGC;
Inner primer 8:ACTGCCTATGCAAACGGCGAGTCCAGGCTGAAGACATTGG.
Embodiment 2: the method that detects Chikungunya virus with a preferred embodiment of test kit of the present invention
Carry out as follows:
Step 1: the pre-treatment of test sample: extract sample rna according to ordinary method.
Step 2: loop-mediated isothermal amplification (LAMP) reaction:
The test kit that present embodiment is selected for use contains the primer of following sequence, the sequence shown in SEQ ID NO:1-4:
Outer primer 1:CGCCCTCTTTAACGGACATG
Outer primer 2:AATTCGGCGCTGGCTAAG
Inner primer 1:TGCCTTTCTTGCTGGCTGCATATACCAGCCTGCACCCATT
Inner primer 2:AGTGTGCGGTGCATTCGATGATGCAGCTGAGAATTCCCTTC.
The preparation reaction solution: reaction system is inner primer 1 and the inner primer 2 of 25uL:1.6mM, the outer primer 1 of 0.2mM and outer primer 2, the dNTPs of 1.6mM, the MgSO4 of 6mM, the trimethyl-glycine of 1M, 2uL sample rna, 8U Bst archaeal dna polymerase, the 2U ThermoScript II, adding distil water is to 25uL.
Above component is mixed back 63 ℃ of insulation 1h.
Step 3: analysis and judgement reaction product result:
Add 1.0 μ l fluorescence dyes (SYBR Green I) in reaction product, mixing leaves standstill 5min.If color is green and then illustrates and have Chikungunya virus in the sample, otherwise then color is orange.
The reaction result of present embodiment is seen Fig. 1, and color is orange behind the tube reaction of Fig. 1 left side, shows that the sample that detects is the Chikungunya virus feminine gender; Color is green behind the tube reaction of right side, shows that the sample that detects is the Chikungunya virus positive.
Select other test kits for use, contain the primer sequence shown in SEQ ID NO:5-8, or the primer sequence shown in SEQ ID NO:9-12, or the primer sequence shown in SEQ ID NO:13-16, by aforesaid detection method last group of test sample detected, can obtain same result.
Above-described embodiment, the present invention a kind of in the embodiment more preferably just, the common variation that those skilled in the art carries out in the technical solution of the present invention scope and replacing all should be included in protection scope of the present invention.
Sequence table
＜120〉a kind of Chikungunya virus isothermal amplification detection kit and primer thereof
<160>16
<170>PatentIn version 3.2
<210>1
<211>20
<212>DNA
＜213〉outer primer 1
<400>1
cgccctcttt aacggacatg 20
<210>1
<211>18
<212>DNA
＜213〉outer primer 2
<400>2
aattcggcgc tggctaag 18
<210>1
<211>40
<212>DNA
＜213〉inner primer 1
<400>3
tgcctttctt gctggctgca tataccagcc tgcacccatt 40
<210>1
<211>41
<212>DNA
＜213〉inner primer 2
<400>4
agtgtgcggt gcattcgatg atgcagctga gaattccctt c 41
<210>1
<211>20
<212>DNA
＜213〉outer primer 3
<400>5
gctgaaaaca cgcagttgag 20
<210>1
<211>20
<212>DNA
＜213〉outer primer 4
<400>6
tggccccaca atgaatttgg 20
<210>1
<211>42
<212>DNA
＜213〉inner primer 3
<400>7
gcggtatgag ccctgtatgc tgaagcacat gtggagaagt cc 42
<210>1
<211>37
<212>DNA
＜213〉inner primer 4
<400>8
cagctaagct ccgcgtcctt tcgccgtttg cataggc 37
<210>1
<211>19
<212>DNA
＜213〉outer primer 5
<400>9
caggcaccat ctggcttta 19
<210>1
<211>20
<212>DNA
＜213〉outer primer 6
<400>10
cccccaaagt ctgaggaatg 20
<210>1
<211>38
<212>DNA
＜213〉inner primer 5
<400>11
gttccctacg gcgcagttca gcagcacaca gcaccatt 38
<210>1
<211>41
<212>DNA
＜213〉inner primer 6
<400>12
gcccatctcc atcgacatac cgcgcacgac atgtccgtta a 41
<210>1
<211>20
<212>DNA
＜213〉outer primer 7
<400>13
cgaagcacat gtggagaagt 20
<210>1
<211>20
<212>DNA
＜213〉outer primer 8
<400>14
agacgtcacc tttgtacacc 20
<210>1
<211>40
<212>DNA
＜213〉inner primer 7
<400>15
ttggtaaagg acgcggagct tttgcatcag catacagggc 40
<210>1
<211>40
<212>DNA
＜213〉inner primer 8
<400>16
actgcctatg caaacggcga gtccaggctg aagacattgg 40
Claims (2)
1. a Chikungunya virus isothermal amplification detection kit is characterized in that, described test kit comprises primer, Bst archaeal dna polymerase, reaction solution and SYBR Green I fluorescence dye;
Described reaction solution is 10mM deoxynucleoside triphosphate, 10 * reaction buffer and 150mM MgSO
4
Described primer sequence is the sequence shown in SEQ ID NO:1-4:
Outer primer 1:CGCCCTCTTTAACGGACATG;
Outer primer 2:AATTCGGCGCTGGCTAAG;
Inner primer 1:TGCCTTTCTTGCTGGCTGCATATACCAGCCTGCACCCATT;
Inner primer 2:AGTGTGCGGTGCATTCGATGATGCAGCTGAGAATTCCCTTC.
2. one kind is used for the primer that the Chikungunya virus isothermal duplication detects, and it is characterized in that described primer sequence is the sequence shown in SEQ ID NO:1-4:
Outer primer 1:CGCCCTCTTTAACGGACATG;
Outer primer 2:AATTCGGCGCTGGCTAAG;
Inner primer 1:TGCCTTTCTTGCTGGCTGCATATACCAGCCTGCACCCATT;
Inner primer 2:AGTGTGCGGTGCATTCGATGATGCAGCTGAGAATTCCCTTC.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011103019932A CN102399903B (en) | 2011-09-30 | 2011-09-30 | Chikungunya virus isothermal amplification detection kit and primer thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011103019932A CN102399903B (en) | 2011-09-30 | 2011-09-30 | Chikungunya virus isothermal amplification detection kit and primer thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102399903A CN102399903A (en) | 2012-04-04 |
| CN102399903B true CN102399903B (en) | 2013-07-03 |
Family
ID=45882546
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2011103019932A Expired - Fee Related CN102399903B (en) | 2011-09-30 | 2011-09-30 | Chikungunya virus isothermal amplification detection kit and primer thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102399903B (en) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105256058B (en) * | 2015-11-20 | 2019-03-26 | 浙江省疾病预防控制中心 | A kind of constant-temperature amplification detection kit and detection method of chikungunya fever virus |
| CN105483293B (en) * | 2016-01-29 | 2019-07-16 | 中国人民解放军疾病预防控制所 | Deadly infectious disease pathogen detection primer sets and kit |
| CN110607401A (en) * | 2019-10-10 | 2019-12-24 | 中国检验检疫科学研究院 | Kit for rapidly detecting chikungunya virus |
-
2011
- 2011-09-30 CN CN2011103019932A patent/CN102399903B/en not_active Expired - Fee Related
Non-Patent Citations (6)
| Title |
|---|
| Clinical Features and Molecular Diagnosis of Chikungunya Fever from South India;Vemu Lakshmi et al.;《Clinical Infectious Diseases》;20080331;1436-1442 * |
| M. M. Parida et al..Rapid and Real-Time Detection of Chikungunya Virus by Reverse Transcription Loop-Mediated Isothermal Amplification Assay.《JOURNAL OF CLINICAL MICROBIOLOGY》.2006,351-357. |
| MM Parida.rapid and real-time assays for detection and quantification of chikunguanya virus.《Future Virol.》.2008,179-192. |
| rapid and real-time assays for detection and quantification of chikunguanya virus;MM Parida;《Future Virol.》;20080331;179-192 * |
| Rapid and Real-Time Detection of Chikungunya Virus by Reverse Transcription Loop-Mediated Isothermal Amplification Assay;M. M. Parida et al.;《JOURNAL OF CLINICAL MICROBIOLOGY》;20061129;351-357 * |
| Vemu Lakshmi et al..Clinical Features and Molecular Diagnosis of Chikungunya Fever from South India.《Clinical Infectious Diseases》.2008,1436-1442. |
Also Published As
| Publication number | Publication date |
|---|---|
| CN102399903A (en) | 2012-04-04 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN110628955B (en) | CrRNA target and CRISPR-Cas13a system for detecting Ebola virus | |
| CN103320434B (en) | Salmonella LAMP (loop-mediated isothermal amplification) primer group and kit and detection method | |
| US8785130B2 (en) | Use of markers including nucleotide sequence based codes to monitor methods of detection and identification of genetic material | |
| CN113322338B (en) | CDA primer group and kit for detecting Shigella and application of CDA primer group and kit | |
| CN102242197B (en) | LAMP (loop-mediated isothermal amplification) kit and special primers thereof for detecting NDM-1 (new Delhi metallo-beta-lactamase 1) gene | |
| CN101736081A (en) | Kit for rapidly detecting isothermal gene amplification of Mycobacterium tuberculosis and detection method | |
| CN102399903B (en) | Chikungunya virus isothermal amplification detection kit and primer thereof | |
| AU2021103978A4 (en) | A primer set, reagent and method based on polymerase spiral reaction for detecting feline parvovirus | |
| Bölske et al. | Diagnosis of paratuberculosis by PCR. | |
| CN117210455B (en) | Primer group, kit and method for detecting east yang virus based on LAMP method | |
| KR102019804B1 (en) | Primer set for detection of SFTSV and SFTSV detection method using the same | |
| EP2597162A1 (en) | Nucleic acid detection | |
| CN101225440A (en) | Detection method of leptospira | |
| Sun et al. | Rapid and sensitive detection of hepatitis C virus in clinical blood samples using reverse transcriptase polymerase spiral reaction | |
| CN113481326B (en) | Isothermal nucleic acid amplification reaction reagent, isothermal nucleic acid amplification method and application thereof | |
| CHENG et al. | Sensitive detection of influenza A (H1N1) virus by isothermal amplification in a single tube | |
| CN110878380B (en) | Primer composition, kit and method for detecting vesicular stomatitis virus Indiana type and new Jersey type | |
| CN114921587A (en) | Nucleic acid detection kit for simultaneously detecting 9 respiratory pathogens based on common qPCR detection platform | |
| CN110093429B (en) | High-flux quantitative detection kit for diarrheal pathogenic bacteria | |
| CN106939356B (en) | Detection primer group, detection kit and detection method for rapidly detecting bee filovirus | |
| RU2455364C2 (en) | Method of identifying mycobacteria by polymerase chain reaction | |
| CN101871015B (en) | Goose parvovirus detection kit and method based on loop-mediated isothermal amplification technology | |
| CN110894549A (en) | RAA constant temperature fluorescence detection method and reagent for Epidemic Hematopoietic Necrosis Virus (EHNV) | |
| CN108950025B (en) | Primer group and kit for rapid detection of mycobacterium tuberculosis katG gene mutation and application | |
| CN110894544A (en) | RAA constant temperature fluorescence detection method and reagent for Salmon Alphavirus (SAV) |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20130703 Termination date: 20140930 |
|
| EXPY | Termination of patent right or utility model |