CN101899534B - Kit and method for detecting goose parvovirus and Muscovy duck parvovirus - Google Patents
Kit and method for detecting goose parvovirus and Muscovy duck parvovirus Download PDFInfo
- Publication number
- CN101899534B CN101899534B CN2010101730821A CN201010173082A CN101899534B CN 101899534 B CN101899534 B CN 101899534B CN 2010101730821 A CN2010101730821 A CN 2010101730821A CN 201010173082 A CN201010173082 A CN 201010173082A CN 101899534 B CN101899534 B CN 101899534B
- Authority
- CN
- China
- Prior art keywords
- parvovirus
- kit
- 10mmol
- muscovy duck
- pcr
- 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.)
- Active
Links
Images
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention relates to a kit for quickly identifying and diagnosing goose parvovirus (GPV) and Muscovy duck parvovirus (MDPV) in an infected Muscovy duck group by using secondary polymerase chain reaction (PCR) technology and a detection method. The kit contains Taq DNA polymerase (5U/mul L), 10*PCR buffer, Mg<2+> (10mmol/L), dNTP (10mmol/L) and two pairs of specific primers. The virus type of the infected Muscovy duck group can be identified by performing extraction and the PCR on DNA in a texture sample, so that the defect that two viruses cannot be quickly identified in the prior art is overcome. The kit has the characteristics of strong specificity, high sensitivity, quickness and the like, and is suitable for clinical quick identification and diagnosis.
Description
Technical field
The invention belongs to biological technical field, relate to the detection kit of a kind of quick discriminating gosling plague virus and kind duck parvovirus, and relate to a kind of method of using this kind test kit to differentiate gosling plague virus and kind duck parvovirus fast.
Background technology
Gosling plague (Gosling Plague GP) claims that again goose parvovirus is sick, find in Yangzhou by fixed 1 years of Chinese scholar side the earliest, and separated with the goose embryo in 1961 should disease virus (Goose Parvovirus, GPV).This disease mainly betides young goose and young kind duck in 1 monthly age, is a kind of height contagious disease fast, that mortality ratio is high of propagating.The young goose M & M of 10 ages in days can reach 100%, and the lethality rate in the susceptible crowd is up to 70%.This disease all has in various degree popular all over the world, extensively betide states such as China, European various countries, Israel, Japan, is one of principal disease of supporting the goose industry of harm at present.
Kind duck parvovirus (Muscovy Duck Parvovirus, MDPV) disease, China's report in 1985, France's report in 1989.M & M was high with interior young bird kind duck (three weeks are sick) 3 ages in week for main infringement, was with enteritis, diarrhoea, weak foot and the acute and subacute infectious disease of breathing and examining characteristic for facing.
The cause of disease goose parvovirus of two kinds of diseases and a kind duck parvovirus all belong to Parvoviridae, parvovirus belongs to.Virus is lattice arrangement under Electronic Speculum, solid and hollow two kinds of particles is arranged, no cyst membrane, diameter 20～24nm; The nucleic acid single stranded DNA, the 5.1kbp size.
Because two kinds of viruses all can infect young kind duck, and symptom is similar with pathological change, can both cause death.Thereby, how two kinds of cause of diseases are differentiated to diagnosing the illness it is an importance fast.
Summary of the invention
The object of the invention just is to develop a kind of detection kit and detection method thereof of differentiating goose parvovirus and kind duck parvovirus.
Technical scheme of the present invention is: a kind of test kit that is used to detect gosling plague virus and kind duck parvovirus comprises following composition:
10 * PCR buffer, Mg2+ (10mmol/L), dNTP (10mmol/L), Taq archaeal dna polymerase (5U/ μ L), 2 pairs of Auele Specific Primers.Wherein:
First pair of primer only is that the gosling plague viral genome is special, and a kind duck parvovirus genome is non-complementary:
F1：5’-gcagg?aacaa?ttacc?ag-3’
R1：5’-acc?acctc?ccgca?ctgac-3’
Second pair of primer is two kinds of fragments that aquatic bird parvovirus genome all has:
F2：5’-cggaa?ggcac?catgc?cgccg-3’
R2：5’-cagaa?tttac?agatt?ttgag-3’
Another technical scheme of the present invention is:
Use the mentioned reagent box to detect the method for gosling plague virus and kind duck parvovirus, its step comprises:
(1) from sample to be tested, extracts viral genome: adopt the DNA extraction test kit to carry out;
(2) carry out PCR simultaneously with two pairs of primers, system is following:
10×pCR?buffer 5μL
Mg2+10mmol/L 3μL
dNTP?10mmol/L 1μL
Taq archaeal dna polymerase 5U/ μ L 1 μ L
Primers F 11 μ L
Primer R1 1 μ L
Primers F 21 μ L
Primer R2 1 μ L
Amplification condition is: 95 ℃ of sex change 5 minutes, 94 ℃ 1 minute, 50 ℃ 1 minute, 72 ℃ 1 minute, 30 circulations, 72 ℃ were extended 10 minutes.
(3) electrophoresis is identified: get 10 microlitre PCR products and carry out electrophoresis with 1% sepharose, if two bands that size is about 774bp and 1145bp size appear in electrophoresis result simultaneously, then be judged to be gosling plague virus and infect; If the band that size is about the 1145bp size only appears in electrophoresis result, then be judged to be a kind duck parvovirus infections; If electrophoresis result does not amplify band, then judging does not have this two kinds of virus infectiones.
The present invention compared with prior art; Its advantage and positively effect show: the present invention has set up the detection kit and the detection method thereof of quick discriminating gosling plague virus and kind duck parvovirus; This test kit according to the design of aquatic bird parvovirus gene order characteristics two pairs of primers; The a pair of distinctive gene fragment of goose parvovirus that can only increase, another is to the two kinds of viral common gene fragments that can increase.The differential diagnosis technology that the present invention sets up, high specificity has very high sensitivity, can be used for the differential diagnosis of gosling plague virus and kind duck parvovirus.
The invention solves and can't differentiate gosling plague virus and kind duck parvovirus defective in the prior art fast, differential diagnosis kit that provides and detection method thereof, fast, accuracy is high, susceptibility good, can be applicable to the veterinary clinic field.
Description of drawings
Fig. 1 is that double PCR identifies GPV and MDPV electrophorogram, and wherein M is DL2000 standard molecular weight Marker, the positive GPV contrast of the 1st swimming lane; The positive MDPV contrast of the 2nd swimming lane; The negative contrast of the 3rd swimming lane, the 4th, 5 swimming lanes are that GPV infects pathological material of disease, and the 6th swimming lane is that MDPV infects pathological material of disease.
Embodiment
Following instance further specifies the present invention, but should not be used as limitation of the present invention.
Embodiment 1: make the detection kit of differentiating gosling plague virus and kind duck parvovirus fast by following prescription:
10 * PCR buffer, Mg2+ (10mmol/L), dNTP (10mmol/L), Taq archaeal dna polymerase (5U/ μ L), 2 pairs of Auele Specific Primers (primer is synthetic by Invitrogen company).Wherein:
First pair of primer only is that the gosling plague viral genome is special, and a kind duck parvovirus genome is non-complementary:
F1：5’-gcagg?aacaa?ttacc?ag-3’(SEQ?ID?NO.1)
R1：5’-acc?acctc?ccgca?ctgac-3’(SEQ?ID?NO.2)
Second pair of primer is two kinds of fragments that aquatic bird parvovirus genome all has:
F2：5’-cggaa?ggcac?catgc?cgccg-3’(SEQ?ID?NO.3)
R2：5’-cagaa?tttac?agatt?ttgag-3’(SEQ?ID?NO.4)
Embodiment 2: utilize the detection kit of differentiating gosling plague virus and kind duck parvovirus fast to detect parvovirus infections situation among kind duck crowd
(1) reaches and extract viral genome infected tissue's (negative control) from pathological material of disease (2 parts of gosling plague virus infection pathological material of diseases, 1 part of kind of duck parvovirus infections pathological material of disease): adopt Quiagen company DNA extraction test kit to carry out;
(2) carry out PCR simultaneously with two pairs of primers, GPV is got in setting simultaneously and MDPV nucleic acid is done positive template contrast, and system is following:
10×PCR?buffer 5μL
Mg2+10mmol/L 3μL
dNTP?10mmol/L 1μL
Taq archaeal dna polymerase 5U/ μ L 1 μ L
Primers F 11 μ L
Primer R1 1 μ L
Primers F 21 μ L
Primer R2 1 μ L
Amplification condition is: 95 ℃ of sex change 5 minutes, 94 ℃ 1 minute, 50 ℃ 1 minute, 72 ℃ 1 minute, 30 circulations, 72 ℃ were extended 10 minutes.
(3) electrophoresis is identified: as shown in Figure 1, gosling plague virus infects pathological material of disease (the 4th, 5 swimming lane) and occurs two bands that size is about 774bp and 1145bp size simultaneously; The band that size is about the 1145bp size only appears in kind duck parvovirus infections pathological material of disease (the 6th swimming lane); And negative control (negative control) does not amplify band.
SEQUENCE?LISTING
< 110>Yangzhou University
< 120>be used to detect the test kit and the detection method thereof of gosling plague virus and kind duck parvovirus
<130>
<160>4
<170>PatentIn?version?3.3
<210>1
<211>17
<212>DNA
< 213>artificial sequence
<400>1
gcaggaacaa?ttaccag 17
<210>2
<211>18
<212>DNA
< 213>artificial sequence
<400>2
accacctccc?gcactgac 18
<210>3
<211>20
<212>DNA
< 213>artificial sequence
<400>3
cggaaggcac?catgccgccg 20
<210>4
<211>20
<212>DNA
< 213>artificial sequence
<400>4
cagaatttac?agattttgag 20
Claims (1)
1. test kit that is used to detect gosling plague virus and kind duck parvovirus comprises following composition:
10 * PCR buffer, Mg
2+10mmol/L, dNTP 10mmol/L, Taq archaeal dna polymerase 5U/ μ L, 2 pairs of Auele Specific Primers, wherein:
First pair of primer is:
F1：?5’-?gcagg?aacaa?ttacc?ag?–3’
R1：5’-?acc?acctc?ccgca?ctgac-3’
Second pair of primer is:
F2：?5’-?cggaa?ggcac?catgc?cgccg?-3’
R2：?5’-?cagaa?tttac?agatt?ttgag?-3’?。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2010101730821A CN101899534B (en) | 2010-05-14 | 2010-05-14 | Kit and method for detecting goose parvovirus and Muscovy duck parvovirus |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2010101730821A CN101899534B (en) | 2010-05-14 | 2010-05-14 | Kit and method for detecting goose parvovirus and Muscovy duck parvovirus |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101899534A CN101899534A (en) | 2010-12-01 |
| CN101899534B true CN101899534B (en) | 2012-07-04 |
Family
ID=43225451
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2010101730821A Active CN101899534B (en) | 2010-05-14 | 2010-05-14 | Kit and method for detecting goose parvovirus and Muscovy duck parvovirus |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101899534B (en) |
Families Citing this family (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102346191B (en) * | 2011-06-17 | 2013-09-11 | 福建省农业科学院畜牧兽医研究所 | Muscovy duck gosling plague latex particle agglutination reagent and preparation method thereof |
| CN102426238B (en) * | 2011-09-07 | 2013-09-04 | 福建省农业科学院畜牧兽医研究所 | Immunofluorescent reagent used for detecting Muscovy duck gosling blast diseases |
| CN102586487B (en) * | 2012-03-15 | 2013-10-02 | 广西壮族自治区兽医研究所 | Double fluorescence quantitative reverse transcription-polymerase chain reaction (RT-PCR) detection kit for duck hepatitis virus I and Muscovy duck parvovirus |
| CN102719564B (en) * | 2012-06-25 | 2013-09-25 | 广西壮族自治区兽医研究所 | Triple polymerase chain reaction (PCR) kit for duck hepatitis virus type I, duck circoviruses and Muscovy duckling parvovirosis and application of triple PCR kit |
| CN103255229B (en) * | 2012-09-05 | 2015-05-27 | 中国农业科学院哈尔滨兽医研究所 | One tube PCR type kit for discriminating and diagnosing goose parvovirus and Muscovy duck parvovirus |
| CN102876808B (en) * | 2012-09-27 | 2014-10-08 | 广西壮族自治区兽医研究所 | Muscovy duckling parvovirus LAMP (loop-mediated isothermal amplification) detection reagent kit |
| CN102876812B (en) * | 2012-10-22 | 2014-01-08 | 金陵科技学院 | Kit for detecting FADV (pigeon adenovirus) |
| CN103409562B (en) * | 2013-09-02 | 2015-03-25 | 福建省农业科学院畜牧兽医研究所 | PCR-RFLP (polymerase chain reaction-restriction fragment length polymorphism) method for distinguishing goose parvovirus from Muscovy duck parvovirus |
| CN109439804A (en) * | 2018-12-20 | 2019-03-08 | 华南农业大学 | The detection primer and detection kit of a kind of Muscovy duck parvovirus and application |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101591703A (en) * | 2008-11-22 | 2009-12-02 | 中国水产科学研究院黄海水产研究所 | The store method of loop-mediated isothermal amplification reaction reagent mixture |
-
2010
- 2010-05-14 CN CN2010101730821A patent/CN101899534B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101591703A (en) * | 2008-11-22 | 2009-12-02 | 中国水产科学研究院黄海水产研究所 | The store method of loop-mediated isothermal amplification reaction reagent mixture |
Non-Patent Citations (3)
| Title |
|---|
| Yun Zhang et al..Development and evaluation of a VP3-ELISA for the detection of goose and Muscovy duck parvovirus antibodies.《Journal of Virological Methods》.2010,第163卷(第2期),405-409. * |
| 张伟等.华南地区小鹅瘟和番鸭"三周病"分离野毒株部分VP基因的序列差异分析.《《黑龙江畜牧兽医》科技版》.2009,(第10期),64-67. |
| 王忠等.小鹅瘟血清学和分子生物学诊断方法.《黑龙江畜牧兽医》.2009,(第1期),88-89. * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101899534A (en) | 2010-12-01 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101899534B (en) | Kit and method for detecting goose parvovirus and Muscovy duck parvovirus | |
| Dowgier et al. | A molecular survey for selected viral enteropathogens revealed a limited role of Canine circovirus in the development of canine acute gastroenteritis | |
| Almasi et al. | Development of colorimetric loop-mediated isothermal amplification assay for rapid detection of the tomato yellow leaf curl virus | |
| CN105603124B (en) | LAMP primer group and detection method for the detection of I subgroup aviadenovirus | |
| CN103409555B (en) | Macrobrachium rosenbergii Nodavirus RT-LAMP-LFD detection method and detection kit thereof | |
| CN113943830A (en) | Primer probe set and kit for simultaneously detecting feline parvovirus, feline herpesvirus type 1 and feline calicivirus | |
| CN103160615A (en) | Multiple PCR primer used for simultaneously detecting infectious Bovine Rhinotracheitis virus and akabane virus as well as its design method | |
| CN103866046A (en) | Detection kit for human herpes viruses EBV and VZV | |
| Kim et al. | Evaluation of three automated nucleic acid extraction systems for identification of respiratory viruses in clinical specimens by multiplex real-time PCR | |
| CN101724712B (en) | Animal insect-borne disease multi-RT-PCR distinguishing and detecting reagent as well as preparation method and application | |
| CN108531651A (en) | A kind of specific primer and its application for detecting and differentiating FAdV-4 and FAdV-8b | |
| US20230250497A1 (en) | One-step nested pcr primers set and kit modified with locked nucleic acid for detecting african swine fever virus | |
| CN101392299A (en) | Equine influenza detection kit and detection method | |
| CN106939357A (en) | H4 hypotypes, H6 hypotypes and the triple RT PCR primer combinations of H9 hypotypes AIV, kit and its application | |
| CN103820574A (en) | Real-time fluorescent quantitation PCR (polymerase chain reaction) parting detection kit for human herpesvirus-6 | |
| CN107937606B (en) | Reagent and method for identifying rabies virus vaccine strain and wild strain | |
| CN103215389B (en) | Porcine reproductive and respiratory syndrome and porcine Japanese B encephalitis dual one-step RT-PCR (Reverse Transcription-Polymerase Chain Reaction) diagnosis kit | |
| CN102329891A (en) | RT-LAMP (reverse transcription-loop-mediated isothermal amplification) primer group for detecting H9 subtype avian influenza virus as well as detection method and application thereof | |
| CN105400908B (en) | A kind of primer, kit and detection method using pyrosequencing techniques detection channel catfish virus | |
| CN104164518A (en) | Real-time fluorescent RT-PCR method for direct detection of influenza A+B viruses | |
| Wang et al. | Visual and label-free ASFV and PCV2 detection by CRISPR-Cas12a combined with G-quadruplex | |
| CN103014180A (en) | Detection primer, probe and detection method of human astrovirus nucleotide | |
| Ji et al. | Simple and visible detection of novel astroviruses causing fatal gout in goslings using one-step reverse transcription polymerase spiral reaction method | |
| CN102108415B (en) | Method and kit for detecting reverse transcription-loop mediated isothermal amplification (RT-LAMP) of pike fry rhabdovirus (RFRV) | |
| CN110846437A (en) | Double PCR (polymerase chain reaction) detection primer group, kit and method for chicken parvovirus and H9 subtype avian influenza virus |
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 | ||
| EE01 | Entry into force of recordation of patent licensing contract |
Application publication date: 20101201 Assignee: Vacbio Co., Ltd. Assignor: Yangzhou University Contract record no.: 2013320000090 Denomination of invention: Kit and method for detecting goose parvovirus and Muscovy duck parvovirus Granted publication date: 20120704 License type: Exclusive License Record date: 20130401 |
|
| LICC | Enforcement, change and cancellation of record of contracts on the licence for exploitation of a patent or utility model |