CN108707693A - A kind of yellow fever virus, chikungunya virus rapid fluorescence PCR detection kit - Google Patents
A kind of yellow fever virus, chikungunya virus rapid fluorescence PCR detection kit Download PDFInfo
- Publication number
- CN108707693A CN108707693A CN201810148229.8A CN201810148229A CN108707693A CN 108707693 A CN108707693 A CN 108707693A CN 201810148229 A CN201810148229 A CN 201810148229A CN 108707693 A CN108707693 A CN 108707693A
- Authority
- CN
- China
- Prior art keywords
- virus
- yellow fever
- fever virus
- pcr
- chikungunya
- 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.)
- Pending
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- 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
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/70—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving virus or bacteriophage
- C12Q1/701—Specific hybridization probes
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- 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
- 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
- C12Q1/686—Polymerase chain reaction [PCR]
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Zoology (AREA)
- Engineering & Computer Science (AREA)
- Wood Science & Technology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Immunology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Health & Medical Sciences (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Biophysics (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- Physics & Mathematics (AREA)
- General Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Genetics & Genomics (AREA)
- Virology (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses a kind of yellow fever virus, chikungunya virus rapid fluorescence PCR detection kit, concrete component is as follows：RT-PCR Buffer, yellow fever virus/internal control primer probe mixed liquor A, chikungunya virus/internal control primer probe mixed liquid B, enzyme system C, positive control, negative control.The present invention has many advantages, such as that detection speed is fast, sensitivity is higher.
Description
Technical field
The present invention relates to biotechnology and medical domain field, specifically a kind of yellow fever virus, chikungunya virus
Rapid fluorescence PCR detection kit.
Background technology
Yellow fever virus（Yellow fever virus, YFV）Belong to flaviviridae, Flavivirus, viral genome is not
Segmented single-stranded positive RNA, only there are one serotypes.Yellow fever（Yellow fever, YF）It is caused by yellow fever virus
Acute infectious disease, mosquito are primary vehicles.Yellow fever is divided to two kinds of urban type and jungle type, in Africa and South America torrid areas
Prevalence is one of 3 kinds of infectious diseases of international hygiene regulations of rules monitoring.The WHO estimations annual YF in the whole world falls ill 200,000, about 30,000
Example is dead.Wherein 90% is happened at Africa.On March 10th, 2016, Beijing was found that China's the first Introduced cases yellow fever cases.But
Never there is the report of local yellow fever example, but not with the direct route course line of the yellow fever Endemic Area such as China and Africa, South America
It disconnecting and leading to, possibility of the yellow fever by sailing through to flight or through other regional transfers immigrations substantially increases, so, yellow fever virus
Quarantine needs quickly sensitive detection means.
Chikungunya virus（Chikungunya fever virus, CHIKV）Category Togaviridae alphavirus, 1997
Year is single strand plus RNA virus, 11805 nucleotide of geneome RNA overall length by definite designation.Chikungunya virus
（Chikungunya fever, CHIK）It is to bite a kind of people, animal of propagation by chikungunya disease poisons yellow-fever mosquito to suffer from acute infection altogether
Disease, Major Epidemic is in Africa and Tropical Asian countries and regions.People is generally susceptible to CHIKV, incubation period usual 2-12d, average
3-7d；Main clinical characteristics are the symptoms such as fever, fash, Nausea and vomiting and severe joint pain.In March, 2008.Examine inspection in Guangdong
Epidemic disease office sanitary inspection laboratory detects CHIKV, is China from going to Sri Lanka to work in labor service personnel's serum sample for coming back home
The first CHIK introduced cases.Since current CHIK is treated without vaccine prevention and specific drug, it is real to carry out doubtful CHIK cases in time
Room diagnosis detection is tested, prevents it from breaking out significant with prevalence.
Currently, domestic and international yellow fever virus（Yellow fever virus, YFV）And chikungunya virus（chikungunya
Fever virus, CHIKV）Diagnostic method, main or PCR by Serologic test, tissue cultures and routine detects,
The methods of real-time fluorescence quantitative PCR.Wherein, Serologic test, tissue cultures, have sensitivity is low, immunological cross-reaction and
The shortcomings of period is long.Conventional PCR needs effectively to be improved there is also the deficiency of easy pollution.Multiple real time fluorescence PCR
Technology is the advanced technology for being combined round pcr and multicolor fluorescence label probe, this method have it is quick, special, sensitive, from
The features such as dynamicization degree is high especially meets extensive, quick diagnosis demand in conjunction with pollution prevention technology processing.
The present invention optimizes primer, probe and amplification system on the basis of real-time fluorescence quantitative PCR, can be in ABI7500
It realizes that one-step RT-PCR detection only needs 45min on fluorescent PCR instrument, one-step RT-PCR is realized in 30min on fast PCR instrument
Augmentation detection.Detection time is greatly shortened through the invention, and the inspection of entry and exit port frontline inspection and quarantine personnel can be improved
Efficiency is surveyed, can not only workload have been reduced but also can quickly obtain detection, to protecting China to defend border safety and preventing case diffusion tool
It is significant.
Currently, the defect that kit generally existing detection sensitivity in the market is low, detection speed is slower, is had
The improvement of effect.
Invention content
Poor, slow defect that the purpose of the present invention is to solve detection sensitivities in the prior art, provides one
Yellow fever virus, chikungunya virus rapid fluorescence PCR detection kit are planted to solve the above problems.
To achieve the goals above, the technical solution of invention is as follows：The invention discloses a kind of yellow fever virus, chikungunyas
Viral rapid fluorescence PCR detection kit, concrete component are as follows：RT-PCR Buffer, yellow fever virus/internal control primer probe are mixed
Close liquid A, chikungunya virus/internal control primer probe mixed liquid B, enzyme system C, positive control, negative control.
Preferably, the invention discloses a kind of yellow fever virus, chikungunya virus rapid fluorescence PCR detection kit,
Concrete component is as follows：384 μ l of RT-PCR Buffer, yellow fever virus/48 μ l of internal control primer probe mixed liquor A, chikungunya disease
Poison/48 μ l of internal control primer probe mixed liquid B, 96 μ l of enzyme system C, 400 μ l of positive control
, 400 μ l of negative control.
Preferably, the group of the RT-PCR Buffer is divided into 50mM tris-HCL, 75mM Kcl, 3mM
The group of Mgcl2,10mM DTT, the yellow fever virus/internal control primer probe mixed liquor A are divided into yellow fever virus and reference gene draws
The group of object, probe, the chikungunya virus/internal control primer probe mixed liquid B is divided into chikungunya virus and reference gene
The group of primer, probe, the enzyme system C is divided into 0.1U/ μ l Hitaq thermal startings archaeal dna polymerase, 2.4U/ μ l super M-MLV
Reverse transcriptase, 0.32U/ μ l RRI, the positive control, which uses, contains yellow fever virus and chikungunya virus target gene piece
The plasmid bacterial of section, the negative control use DEPC-H2O.
A kind of above-mentioned yellow fever virus, chikungunya virus rapid fluorescence PCR detection kit detection yellow fever virus, datum hole are agreed
The primer probe sequence of refined virus, specifying information are as follows：
Preferably, the yellow fever virus, chikungunya virus probe 5 ' hold FAM, JOE, FITC fluorescein, 3 ' end labels
BHQ1, TAMRA or BHQ2 quench fluorescein, and reference gene 5 ' holds HEX, ROX or CY5 mark fluorescent plain, 3 ' end label BHQ1,
TAMRA or BHQ2 quenches fluorescein.
It is a kind of that yellow fever virus, base are detected using above-mentioned yellow fever virus, chikungunya virus rapid fluorescence PCR detection kit
The Fluorescence PCR system of refined virus is agreed in hole, specific as follows：
It is a kind of to utilize above-mentioned yellow fever virus, chikungunya virus rapid fluorescence PCR detection kit to yellow fever virus, datum hole
Agree refined viral fast PCR detection, the detection method comprises the steps of：
（1）, carry out sample rna extraction
With the QIAampMinElute Virus Spin Kit of QIAGEN companies（Article No.：57704）Or Tiangeng biochemical technology is limited
Company's viral RNA extracts kit（Article No.：DP315）Or the viral RNA extracts reagent of Anhui Anlong Gene Tech. Company Limited
Box（Article No.：AL-L0107）, sample rna is extracted；
（2）, PCR reaction systems preparation
PCR reaction systems are prepared in preparation of reagents room, 16 μ l of PCR reaction buffers, 2 μ l of enzyme system C, primed probe
PCR reaction solution is dispensed by 20 μ l/ pipes into PCR reaction tubes, the reaction tube equipped with PCR reaction solution is moved to by 2 μ l of mixed liquor
Sample process area；
（3）, sample-adding
Each 5 μ l of processed sample, negative control, positive control supernatant are taken to be added separately to fill respectively with the suction nozzle with filter core
Have in the PCR reaction tubes of reaction system, lid upper tube cap, augmentation detection area is moved to after centrifuging the several seconds.
（4）, amplification
PCR reaction tubes are put into fluorescent PCR amplification instrument and carry out augmentation detection
Preferably, the step（4）In, loop parameter setting is as follows：（AB：ABI 7500FAST, Anlong：Pro
Eco48, SLAN-96）：
The present invention has the following advantages compared with prior art：
1, high sensitivity：Virus of the detectable template concentrations down to 100PFU/ml；
2, high specificity, the detection architecture monitored using reference gene, ensures the reliability of testing result；
3, precision is high, and kit repeats detection 10 times, intermediate value cv< to intermediate value and low value precision standard items;1%, low value cv<
2%；
4, detection time is short, and the PCR reaction Buffer and enzyme system prepared using kit 45min on fast PCR instrument are realized
Reverse transcription and 40 cyclic amplification detections, can be improved the detection efficiency of entry and exit port frontline inspection and quarantine personnel, can both subtract
Few workload can quickly obtain detection again, to protecting China to defend border safety and case diffusion being prevented to be of great significance.
Description of the drawings
The yellow fever of yellow fever virus, chikungunya virus rapid fluorescence PCR detection kit that Fig. 1 is prepared for embodiment 1
Malicious detection sensitivity result figure；
Fig. 2 is that yellow fever virus, the datum hole of chikungunya virus rapid fluorescence PCR detection kit that embodiment 1 is prepared are willing
Refined viral diagnosis sensitivity results figure；
Fig. 3 is that yellow fever virus, chikungunya virus rapid fluorescence PCR detection kit detection Huang that embodiment 1 is prepared are warm
Viral precision result figure；
Fig. 4 is yellow fever virus, the chikungunya virus rapid fluorescence PCR detection kit detection datum hole that embodiment 1 is prepared
Agree refined viral density result figure.
Specific implementation mode
To make the structure feature to invention and the effect of being reached has a better understanding and awareness, to preferably real
Example and attached drawing cooperation detailed description are applied, is described as follows：
Referring to Fig.1-4,
Embodiment 1
One, nucleic acid extraction is carried out in sample process area
1.1, use the QIAampMinElute Virus Spin Kit of QIAGEN companies（Article No.：57704）, Tiangeng biochemistry section
Skill Co., Ltd virus genom DNA/RNA extracts kits（Article No.：DP315）Nucleic acid is extracted；
1.2, positive control and negative control and sample to be tested synchronize handled；
Two, PCR reagent preparation is carried out in reagent area in preparation
2.1, it presses following composition and prepares PCR reaction solution（N is reaction tube number）
PCR reaction buffers 1 are 16 μ l × n, and yellow fever virus primer/internal standard probe mixed liquor 1 is 2 μ l × n, chikungunya disease
Malicious primer/internal standard probe mixed liquor 2 is 2 μ l × n, and enzyme system 3 is 2 μ l × n, and PCR reaction buffers, primer are ensured before use
Probe mixed liquor fully dissolves, and enzyme system needs centrifugation to ensure that all enzymes concentrate on bottom before use；
2.2, PCR reaction solutions are dispensed by 20 μ l/ pipes into PCR reaction tubes, the reaction tube equipped with PCR reaction solution is moved into sample
Present treatment area.
Three, it is loaded in sample process area
Each 5 μ l of processed sample, negative control, positive control supernatant are taken to be added separately to be equipped with respectively with the suction nozzle with filter core
In the PCR reaction tubes of PCR reaction solution, lid upper tube cap moves to augmentation detection area after centrifuging the several seconds；
Four, PCR amplification detection is carried out in augmentation detection area
4.1, PCR reaction tubes are put into fluorescent PCR amplification instrument and carry out augmentation detection.
4.2, loop parameter is set：（ABI 7500, SLAN96P）：
5、
6, testing result is explained
（1）Negative findings judge：If the channels sample FAM amplification curve is not S-type, Ct values are UNDET, the channels internal reference HEX are expanded
Increase that curve is S-type and result Ct values <35, then result is feminine gender；If the channels internal reference HEX amplification curve is not S-type or result Ct values
>=35, then sample should be rechecked.
（2）Positive findings judge：If the channels sample FAM amplification curve is S-type and value≤38 Ct, result are the positive.
（3）Test gray area：If the channels sample FAM amplification curve is S-type and result 38<Ct<40, then result is positioned at real
Gray area is tested, sample should be rechecked, if reinspection result amplification curve is S-type, judges result for the positive, if amplification curve
It is not S-type, then judge result for feminine gender.
Embodiment 2
With in yellow fever virus/chikungunya virus RNA detection kits, low two concentration（1×104PFU/ml、1×
103PFU/ml）Yellow fever virus/chikungunya virus as sample to be checked, with yellow fever virus/chikungunya disease of quality inspection qualification
Malicious RNA detection kits are detected, each 8 times repetitions detect.It is detected in ABI7500 instruments.Calculate testing result Ct
It is worth the coefficient of variation.Batch interior Ct value coefficient of variation≤2% of positive sample.
Embodiment 3
Selection and yellow fever virus and chikungunya virus sequence with homology, easily cause same or analogous clinical symptoms, adopt
The normal parasitic or easily concurrent other pathogens in sample position, such as dengue fever virus, zika virus, Lassa virus, Hantaan virus, season
Section property H1N1 influenza viruses, seasonality H3N2 influenza viruses, influenza B virus are sample to be checked, with quality inspection qualification
20140501,20140502,20,140,503 3 batches of yellow fever virus/chikungunya virus kit for detecting nucleic acid are detected, behaviour
Work is carried out in strict accordance with kit specification, is detected on ABI7500 real-time fluorescence quantitative PCR instruments, testing result is equal
For feminine gender, show that kit specificity is good
The advantages of basic principles and main features and the invention of invention has been shown and described above.The technical staff of the industry should
Understand, invention is not restricted to the described embodiments, and what is described in the above embodiment and the description is only the principles of invention, are not taking off
Invention will also have various changes and improvements under the premise of from spirit and range, these changes and improvements both fall within claimed
Invention in the range of.The protection domain that invention requires is defined by appended claims and its equivalent.
Sequence table
<110>Lianyun Harbour Entry-Exit Inspection and Quarantine Bureau
<120>A kind of yellow fever virus, chikungunya virus rapid fluorescence PCR detection kit
<141> 2018-02-13
<160> 9
<170> SIPOSequenceListing 1.0
<210> 1
<211> 20
<212> DNA
<213>Artificial sequence (artificial sequences)
<220>
Claims (8)
1. a kind of yellow fever virus, chikungunya virus rapid fluorescence PCR detection kit, it is characterised in that：Concrete component is as follows：
RT-PCR Buffer, yellow fever virus/internal control primer probe mixed liquor A, chikungunya virus/internal control primer probe mixed liquid B,
Enzyme system C, positive control, negative control.
2. a kind of yellow fever virus according to claim 1, chikungunya virus rapid fluorescence PCR detection kit, special
Sign is：Concrete component is as follows：384 μ l of RT-PCR Buffer, yellow fever virus/48 μ l of internal control primer probe mixed liquor A, datum hole
Agree refined virus/48 μ l of internal control primer probe mixed liquid B, 96 μ l of enzyme system C, 400 μ l of positive control, 400 μ l of negative control.
3. a kind of yellow fever virus according to claim 1, chikungunya virus rapid fluorescence PCR detection kit, special
Sign is：The group of the RT-PCR Buffer is divided into 50mM tris-HCL, 75mM Kcl, 3mM Mgcl2,10mM DTT,
The group of the yellow fever virus/internal control primer probe mixed liquor A is divided into yellow fever virus and reference gene primer, probe, described
The group of chikungunya virus/internal control primer probe mixed liquid B is divided into chikungunya virus and reference gene primer, probe, described
Enzyme system C group be divided into 0.1U/ μ l Hitaq thermal startings archaeal dna polymerase, 2.4U/ μ l super M-MLV reverse transcriptase,
0.32U/ μ l RRI, the positive control use the plasmid containing yellow fever virus and chikungunya virus target gene fragment
Bacterium, the negative control use DEPC-H2O.
4. a kind of yellow fever virus as described in any one of claims 1-3, chikungunya virus rapid fluorescence PCR detection kit are examined
The primer probe sequence of survey yellow fever virus, chikungunya virus, it is characterised in that：Specifying information is as follows：
5. the primer probe sequence of a kind of detection yellow fever virus according to claim 4, chikungunya virus, feature exist
In：The yellow fever virus, chikungunya virus probe 5 ' hold FAM, JOE, FITC fluorescein, 3 ' end label BHQ1, TAMRA or
BHQ2 quenches fluorescein, and reference gene 5 ' holds HEX, ROX or CY5 mark fluorescent element, 3 ' end label BHQ1, TAMRA or BHQ2 sudden
Go out fluorescein.
6. a kind of using yellow fever virus as described in any one of claims 1-3, chikungunya virus rapid fluorescence PCR detection reagents
The Fluorescence PCR system of box detection yellow fever virus, chikungunya virus, it is characterised in that：It is specific as follows：
7. a kind of utilizing yellow fever virus as described in any one of claims 1-3, chikungunya virus rapid fluorescence PCR detection reagents
Box detects yellow fever virus, chikungunya virus fast PCR, it is characterised in that：The detection method comprises the steps of：
(1), the extraction of sample rna is carried out
With the QIAampMinElute Virus Spin Kit (article No.s of QIAGEN companies：57704) or Tiangeng biochemical technology is limited
Company's viral RNA extracts kit (article No.：) or the viral RNA extracts reagent of Anhui Anlong Gene Tech. Company Limited DP315
Box (article No.：AL-L0107), sample rna is extracted；
(2), the preparation of PCR reaction systems
PCR reaction systems are prepared in preparation of reagents room, PCR reaction buffers 16 μ l, 2 μ l of enzyme system C, primed probe are mixed
2 μ l of liquid are closed, PCR reaction solution is dispensed by 20 μ l/ pipes into PCR reaction tubes, the reaction tube equipped with PCR reaction solution is moved into sample
Treatment region；
(3), it is loaded
Each 5 μ l of processed sample, negative control, positive control supernatant are taken to be added separately to be equipped with respectively with the suction nozzle with filter core
In the PCR reaction tubes of reaction system, lid upper tube cap moves to augmentation detection area after centrifuging the several seconds.
(4), it expands
PCR reaction tubes are put into fluorescent PCR amplification instrument and carry out augmentation detection.
8. detection method according to claim 7, it is characterised in that：In the step (4), loop parameter is set such as
Under：(AB：ABI 7500FAST, Anlong：Pro Eco48, SLAN-96)：
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810148229.8A CN108707693A (en) | 2018-02-13 | 2018-02-13 | A kind of yellow fever virus, chikungunya virus rapid fluorescence PCR detection kit |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810148229.8A CN108707693A (en) | 2018-02-13 | 2018-02-13 | A kind of yellow fever virus, chikungunya virus rapid fluorescence PCR detection kit |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN108707693A true CN108707693A (en) | 2018-10-26 |
Family
ID=63866389
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810148229.8A Pending CN108707693A (en) | 2018-02-13 | 2018-02-13 | A kind of yellow fever virus, chikungunya virus rapid fluorescence PCR detection kit |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108707693A (en) |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102776297A (en) * | 2012-08-09 | 2012-11-14 | 中山大学达安基因股份有限公司 | Reagent kit for distinguishing and detecting dengue virus/yellow fever virus/west nile virus/chikungunya virus |
| WO2013080307A1 (en) * | 2011-11-29 | 2013-06-06 | 株式会社 東芝 | Primer set for amplifying mosquito-borne virus, assay kit for detecting mosquito-borne virus, and detection method using said primer set and said assay kit |
| CN103305633A (en) * | 2013-06-14 | 2013-09-18 | 浙江国际旅行卫生保健中心 | Multiplex-fluorescence PCR (Polymerase Chain Reaction) detection kit and application thereof |
| CN106086242A (en) * | 2016-07-29 | 2016-11-09 | 广州市第八人民医院 | A kind of test kit detected for Flavivirus fast typing and virus load |
| CN106755573A (en) * | 2016-12-07 | 2017-05-31 | 深圳澳东检验检测科技有限公司 | Zika virus, dengue fever virus, the RT PCR detection methods of chikungunya fever virus, primer and probe and kit |
-
2018
- 2018-02-13 CN CN201810148229.8A patent/CN108707693A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2013080307A1 (en) * | 2011-11-29 | 2013-06-06 | 株式会社 東芝 | Primer set for amplifying mosquito-borne virus, assay kit for detecting mosquito-borne virus, and detection method using said primer set and said assay kit |
| CN102776297A (en) * | 2012-08-09 | 2012-11-14 | 中山大学达安基因股份有限公司 | Reagent kit for distinguishing and detecting dengue virus/yellow fever virus/west nile virus/chikungunya virus |
| CN103305633A (en) * | 2013-06-14 | 2013-09-18 | 浙江国际旅行卫生保健中心 | Multiplex-fluorescence PCR (Polymerase Chain Reaction) detection kit and application thereof |
| CN106086242A (en) * | 2016-07-29 | 2016-11-09 | 广州市第八人民医院 | A kind of test kit detected for Flavivirus fast typing and virus load |
| CN106755573A (en) * | 2016-12-07 | 2017-05-31 | 深圳澳东检验检测科技有限公司 | Zika virus, dengue fever virus, the RT PCR detection methods of chikungunya fever virus, primer and probe and kit |
Non-Patent Citations (1)
| Title |
|---|
| 郑夔等: "应用含内参的多重实时荧光RT-PCR方法快速检测登革病毒和基孔肯雅病毒", 《中国人兽共患病学报》 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Nagaraj et al. | Ante mortem diagnosis of human rabies using saliva samples: comparison of real time and conventional RT-PCR techniques | |
| CN112063756B (en) | Method and kit for multiple detection of respiratory virus nucleic acid | |
| CN108330210B (en) | Zika virus, dengue virus and chikungunya virus nucleic acid detection kit and application thereof | |
| CN103074449B (en) | Kit for synchronously detecting thirteen diarrhea viruses and detection method of kit | |
| CN106167833B (en) | A kind of while 8 kinds of entomophila encephalitis viruses of detection RT-PCR primers and probe combinations and kit | |
| CN107475446A (en) | Multi-PCR detection method and its probe groups and kit a kind of while that detect various respiratory road virus | |
| CN106811550A (en) | A kind of type vaccine strain of GCRV II and street strain's diagnostic primerses and kit and diagnosis detecting method containing it | |
| CN108034764A (en) | Multiplex PCR detection Coxsackie virus, enterovirns type 71 and enterovirus universal primed probe group | |
| CN107365862A (en) | For detecting the primer and probe and its kit of Echinococcus granulosus in dog excrement | |
| CN108676913A (en) | A kind of human parainfluenza viruses' nucleic acid is hands-free to take gene parting detecting reagent | |
| WO2021190633A1 (en) | Primer for detecting sars-cov-2 novel coronavirus, and test kit, detection method, and application thereof | |
| CN105936946A (en) | One step method inverse transcription PCR kit for detecting and differentiating Zika viruses and detection method thereof | |
| CN105713993B (en) | For detecting PCR primer group, probe groups and the kit of various respiratory road virus | |
| CN106148506A (en) | The antenatal detection kit of a kind of B race streptococcus quantitative fluorescent PCR and application thereof | |
| CN107619885A (en) | A kind of fluorescence RT RAA primers, probe and detection method for being used to detect dengue virus | |
| CN103255232B (en) | Dual fluorescence RT (Reverse Transcription)-PCR (Polymerase Chain Reaction) detection kit and method for avian influenza H7N9 virus | |
| WO2022257002A1 (en) | Rt-pcr detection reagent for detecting novel coronavirus, kit and detection method thereof | |
| CN105483289B (en) | A kind of detection kit and reaction system of the triple direct fluorescence RT-PCRs of enterovirus | |
| CN110343784A (en) | The composition and kit of quadruple influenza nucleic acids detection based on melting curve | |
| CN103409554B (en) | Nucleic acid detection kit for rapidly detecting measles virus/rubella virus | |
| CN108531647A (en) | A kind of zika virus one-step method fluorescence rt-PCR detection methods and kit | |
| CN116024386B (en) | Primer probe combination and kit for detecting novel coronaviruses and distinguishing Omicron different mutant strains | |
| CN101392299A (en) | Equine influenza detection kit and detection method | |
| CN110066889A (en) | Quick detection primer group, kit and its application of the GeXP of four kinds of Flavivirus virus are detected simultaneously | |
| CN108707693A (en) | A kind of yellow fever virus, chikungunya virus rapid fluorescence PCR detection kit |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20181026 |
|
| RJ01 | Rejection of invention patent application after publication |