CN105368988A - Primer group and kit for H10N8 subtype AIV (avian influenza virus) double-RT-PCR (reverse transcription-polymerase chain reaction) detection and applications of primer group and kit - Google Patents
Primer group and kit for H10N8 subtype AIV (avian influenza virus) double-RT-PCR (reverse transcription-polymerase chain reaction) detection and applications of primer group and kit Download PDFInfo
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Abstract
The invention belongs to the technical field of poultry virus detection and discloses a primer group and a kit for H10N8 subtype AIV (avian influenza virus) double-RT-PCR (reverse transcription-polymerase chain reaction) detection and applications of the detection primer group and the kit. The primer group for the H10N8 subtype AIV double-RT-PCR detection comprises two pairs of specific primers, and the two pairs of specific primers include a primer pair H10-1 and H10-2 and a primer pair N8-1 and N8-2 and have sequences represented as SEQ ID No.1-SEQ ID No.4 respectively. The detection kit capable of identifying H10N8 subtype AIVs is produced based on a double-RT-PCR method. The H10N8 subtype AIV double-RT-PCR kit has high specificity and sensitivity and can detect H10 and N8 subtype AIVs simultaneously through one tube, and the simple, fast and effective detection kit is provided for H10N8 subtype AIV detection and has great significance for epidemiological investigation and differential diagnosis of the H10N8 subtype AIVs.
Description
Technical field
The invention belongs to avian viral detection technique field, particularly relate to a kind of H10N8 subtype avian influenza virus duplex RT-PCR and detect primer sets, test kit, and qualification H10 subtype avian influenza virus, the primer pair of N8 subtype avian influenza virus or primer pair group and test kit thereof.
Background technology
Avian influenza virus (AvianInfluenzaVirus, AIV) according to surface glycoprotein (Hemagglutinin, and neuraminidase (Neuraminidase HA), NA) antigenic specificity is divided into different hypotypes, has found that there is 18 kinds of HA hypotypes (H1-H18) and 11 kinds of NA hypotypes (N1-N11) at present.AIV a kind ofly has coating, sub-thread minus strand, segmented RNA viruses, and viral genome is made up of 8 segmented strand RNAs.When the genomic segmented characteristic of AIV makes two or more different subtype influenza virus co-infection cell, progeny virus its nucleic acid fragment when assembling may be reset with the form producer of Homo~logous exchange and cause antigenic shift, thus generation can cause pandemic new epidemic strain.
On December 6th, 2013, the 73 years old woman in one, Nanchang City, Jiangxi Province suffers from H10N8 subtype avian influenza, and because of respiratory insufficiency, shock death, this is the Lethal cases of global the first human infection H10N8.On January 25th, 2014 is also in Nanchang, makes a definite diagnosis again a routine H10N8 subtype avian influenza.
At present, method and the exploitation corresponding reagent of setting up a kind of rapid detection H10N8 avian influenza virus is badly in need of.
Summary of the invention
The technical problem to be solved in the present invention is to provide a kind of H10N8 subtype avian influenza virus duplex RT-PCR and detects primer sets, test kit and application thereof.
For solving the problems of the technologies described above, the present invention by the following technical solutions:
For the identification of a primer sets for H10N8 subtype avian influenza virus, comprise 2 pairs of Auele Specific Primers, be primer pair H10-1 and H10-2, primer pair N8-1 and N8-2 respectively, it has the sequence as shown in SEQIDNo.1 to SEQIDNo.4 respectively.
For the identification of a primer pair for H10 subtype avian influenza virus, comprise Auele Specific Primer H10-1 and H10-2, they have the base sequence of sequence table SEQ IDNo.1 to SEQIDNo.2 respectively.
For the identification of a primer pair for N8 subtype avian influenza virus, comprise Auele Specific Primer N8-1 and N8-2, they have the base sequence of sequence table SEQ IDNo.3 to SEQIDNo.4 respectively.
A kind of H10N8 subtype avian influenza virus duplex RT-PCR detection kit, comprise Reverse Transcription, PCR reagent, DEPC water and the ultrapure water without RNA enzyme, containing 2 pairs of Auele Specific Primers in described PCR reagent, it has the sequence as shown in SEQIDNo.1 to SEQIDNo.4 respectively.
As preferably, described Auele Specific Primer H10-1 and H10-2, N8-1 and the N8-2 final concentration in described PCR reagent is 0.1-0.8 μM.
As preferably, described Reverse Transcription is pipe 16 μ L often, wherein 5 × ReverseTranscriptaseBuffer10 μ L, 50pmol9merRandomPrimer2 μ L, 10mMdNTPMixture2 μ L, 40URibonucleaseInhibitor1 μ L, 5U/ μ LAMVReverseTranscriptase1 μ L.
As preferably, the volume of described PCR reagent is that 26.2 μ L/ manage, and wherein comprises 2 × EasyTaqTMPCRSuperMix25 μ L, 50 μMs of each 0.2 μ L of H10-1 and H10-2 primer, 50 μMs of each 0.4 μ L of N8-1 and N8-2 primer.
Present invention also offers the described primer sets for the identification of H10N8 subtype avian influenza virus, described primer pair or described H10N8 subtype avian influenza virus duplex RT-PCR detection kit and differentiate the application in H10N8 subtype avian influenza virus, H10 subtype avian influenza virus and N8 subtype avian influenza virus.
The present invention has following beneficial effect:
1. contriver is specially for H10 HA Gene of H 9 Subtype AIV, N8 subtype avian influenza virus NA gene design 2 pairs of Auele Specific Primers, be combined into H10N8 subtype avian influenza virus duplex RT-PCR detection kit, a pipe detects the infection that can determine whether as H10N8 subtype avian influenza virus, H10 subtype avian influenza virus and N8 subtype avian influenza virus.Utilize primer pair of the present invention, primer pair group or test kit, can detect in testing sample and whether infect containing H10N8 subtype avian influenza virus, H10 subtype avian influenza virus and N8 subtype avian influenza virus.The present invention establishes the duplex RT-PCR detection kit that a RT-PCR reaction just can detect simultaneously and differentiate H10 subtype avian influenza virus, N8 subtype avian influenza virus, H10N8 subtype avian influenza virus.By amplified conditions optimization, specificity and sensitivity test, set up the duplex RT-PCR detection kit of H10 subtype avian influenza virus and N8 subtype avian influenza virus, this test kit has high specificity, highly sensitive, the advantage such as low cost, high-level efficiency, can be used for clinical detection H10N8 subtype avian influenza virus.
2., because the present invention utilizes the difference of expanding fragment length directly to judge amplification in design of primers, make this method easier when result judges, directly perceived and practical.Experiment proves, this method increases to the H10 subtype avian influenza virus in same sample and N8 subtype avian influenza virus template, the 267bp (H10 subtype avian influenza virus) that result all obtains conforming to test design, the specific amplification band of 464bp (N8 subtype avian influenza virus) are negative to the detection of the common poultry diease of newcastle disease, infectious bronchitis and infectious laryngotracheitis entirely.Sensitivity test result shows, the most low energy of this method detects 10
3the H10 subtype avian influenza virus of copy/μ L, N8 subtype avian influenza virus.
3. the present invention can be H10 hypotype, N8 hypotype, H10N8 subtype avian influenza virus morbidity diagnostic result is accurately provided in early days, significant to its route of transmission of cut-out, can isolate in time and process, significant to effective prevention and control of H10N8 subtype avian influenza, have a extensive future, also have important realistic meaning to aviculture Sustainable development.
Accompanying drawing explanation
Fig. 1 sets up by the present invention the specific test result figure of H10N8 hypotype AIV duplex RT-PCR detection kit;
Wherein: M-DL1000Marker; 1-H10+N8; 2-H10N7; 3-H6N8; 4-H1N1; 5-H2N3; 6-H3N2; 7-H4N6; 8-H5N1; 9-H7N2; 10-H8N4; 11-H9N2; 12-H11; 13-H12N5; 14-H13N6; 15-H14N5; 16-H15N9; 17-NDV; 18-IBV; 19-ILTV; 20-blank.
Fig. 2 sets up by the present invention the sensitivity test result figure of H10N8 hypotype AIV duplex RT-PCR detection kit;
Wherein: M-DL1000Marker; 1-10
8copy/μ L; 2-10
7copy/μ L; 3-10
6copy/μ L; 4-10
5copy/μ L; 5-10
4copy/μ L; 6-10
3copy/μ L; 7-100 copy/μ L; 8-10 copy/μ L; 9-blank.
Embodiment
Below in conjunction with specific embodiment, elaboration detailed is further done to the present invention, but embodiments of the present invention are not limited to the scope that embodiment represents.These embodiments only for illustration of the present invention, but not for limiting the scope of the invention.In addition, after reading content of the present invention, those skilled in the art can do various amendment to the present invention, and these equivalent variations fall within appended claims limited range of the present invention equally.
The experimental technique used in following embodiment if no special instructions, is ordinary method.The material used in following embodiment, reagent etc., if no special instructions, all can obtain from commercial channels.
AIV strain (H3N8, H6N8, H1N1, H3N2, H4N6, H9N2 and H11), NDV, IBV and ILTV are preserved by Guangxi veterinary institute isolation identification; AIV strain (H2N3, H8N4, H10N3, H12N5) is so kind as to give by Hong Kong University; The RNA (H10N7, H5N1, H7N2, H13N6, H14N5 and H15N9) of AIV strain is so kind as to give by Connecticut, USA university.MinibestviralRNA/DNAextractionkitver.4.0 and Reverse Transcription are purchased from the precious biotech firm in Dalian; Glue reclaims test kit purchased from Axygen company; PCRSuperMix is purchased from Beijing Quan Shi gold biotech firm.
embodiment 1: the preparation of avian influenza virus duplex RT-PCR design of primers and test kit
(1) design of primer
According to H10, N8 hypotype AIVHA in GenBank and NA gene, conservative region is found out by DNAStar software aligned sequences, with the amplification condition of Primer5.0 software analysis primer, verified by NCBIBlast, 2 pairs of Auele Specific Primers (see table 1) are designed and synthesized, for the detection of H10 subtype avian influenza virus and N8 subtype avian influenza virus.
Table 1 primer information
Note: AIVH10-1 and H10-2 primer pair is used for detecting whether contain H10 subtype avian influenza virus; AIVN8-1 and N8-2 primer pair is used for detecting whether contain N8 subtype avian influenza virus.
(2) preparation of sample
Extract test kit specification sheets with reference to MinibestviralRNA/DNAextractionkitver.4.0, extract the RNA of above-mentioned avian influenza virus, Avian pneumo-encephalitis virus and infectious bronchitis virus respectively, extract the DNA of infectious laryngotracheitis virus.
(3) preparation of H10N8 avian influenza virus duplex RT-PCR detection kit
RT-PCR primer concentration, temperature of reaction, time and cycle index etc. are optimized, finally determine that the best effort final concentration of H10-1 and H10-2 primer in RT-PCR is 0.2 μM; N8-1 and N8-2, primer best effort final concentration be 0.4 μM; The optimum response pattern of PCR is 94 DEG C of denaturation 5min, then enters 94 DEG C of sex change 30s, 52 DEG C of annealing 30s, and 72 DEG C extend 40s, carry out 35 circulations altogether, after finally extending 10min through 72 DEG C again, terminate reaction in 10 DEG C.
Reverse Transcription (purchased from the precious biotech firm in Dalian) comprises 5 × ReverseTranscriptaseBuffer, 50pmol9merRandomPrimer, 10mMdNTPMixture, 40URibonucleaseInhibitor, 5U/ μ LAMVReverseTranscriptase and DEPC water.
Reverse Transcription concrete often pipe 16 μ L: wherein 5 × ReverseTranscriptaseBuffer10 μ L, 50pmol9merRandomPrimer2 μ L, 10mMdNTPMixture2 μ L, 40URibonucleaseInhibitor1 μ L, 5U/ μ LAMVReverseTranscriptase1 μ L.When carrying out reverse transcription, add the RNA template that 1pg-1 μ g is to be checked, DEPC water complements to 50 μ L, and its reverse transcription is become cDNA.
The optimum response pattern of reverse transcription is 25 DEG C of 10min, then 42 DEG C of 50min, last 95 DEG C of 2min.
Adopted by primer pair sterilizing ultrapure water to dissolve, upstream and downstream primer concentration is 50 μMs.
PCR reagent comprises 2 × EasyTaq
tMpCRSuperMix (purchased from Beijing Quan Shi gold biotech firm), primer pair H10-1 and H10-2, primer pair N8-1 and N8-2; The volume of PCR reagent is that 26.2 μ L/ manage, and wherein comprises 2 × EasyTaq
tMpCRSuperMix25 μ L, 50 μMs of each 0.2 μ L of H10-1 and H10-2 primer, 25 μMs of each 0.4 μ L of N8-1 and N8-2 primer.
When detecting for H10N8 hypotype AIV duplex RT-PCR, in PCR reagent, add cDNA2 μ L to be checked, to supply 50 μ L without the ultrapure water of RNA enzyme.
embodiment2: the specific detection of duplex RT-PCR test kit
According to the reaction conditions optimized, detect with the RNA/DNA of set up H10N8 hypotype AIV duplex RT-PCR detection kit to H1N1, H2N3, H3N2, H4N6, H5N1, H6N8, H7N2, H8N4, H9N2, H10N3, H10N7, H11, H12N5, H13N5, H14N5, H15N9, NDV, IBV and ILTV, check its specificity.
As shown in Figure 1, use this test kit to occur 2 specific bands to H10+N8 hypotype AIV detected result, be respectively 267bp and 464bp; Only occur 1 specific band to H10 hypotype AIV (i.e. H10Ny hypotype AIV, y ≠ 8) detected result, clip size is 267bp; Only occur 1 specific band to N8 hypotype AIV (i.e. HxN8 hypotype AIV, x ≠ 10) detected result, clip size is 464bp; Any band is not all amplified to other hypotype AIV and common poultry diease pathogenic agent, conforms to desired design.
embodiment3: the sensitivity test of duplex RT-PCR detection kit
With HA and NA total length primer, respectively with the RNA of H10 hypotype and N8 hypotype AIV for template carries out RT-PCR amplification, obtain the object fragment of full length gene, these two gene fragments are connected on pGEM-TEasy carrier respectively, by the recombinant plasmid of the correct sequence containing HA and NA gene fragment called after T-H10 and T-N8 respectively, respectively by its 10 times of gradient dilutions, according to the reaction conditions optimized, with set up test kit, pcr amplification is carried out to it, check its susceptibility.
As shown in Figure 2, this test kit is 10 to concentration
8the recombinant plasmid of T-H10 and T-N8 of-10 copy/μ L carries out sensitivity testing respectively, and result shows, and is 10 to concentration
8-10
3h10, N8 hypotype AIV of copy/μ L all has 2 obvious amplified bands to occur, clip size is respectively 267bp and 464bp; Be that the H10N8 hypotype AIV of 100 copies/μ L and 10 copy/μ L is all without amplified band to concentration.Therefore, the most low energy of this test kit detects 10
3the H10N8 hypotype AIV of copy/μ L.
Above result shows that utilizing this duplex RT-PCR test kit to detect H10N8 hypotype AIV has higher sensitivity.
Claims (8)
1. for the identification of a primer sets for H10N8 subtype avian influenza virus, it is characterized in that: comprise 2 pairs of Auele Specific Primers, be primer pair H10-1 and H10-2, primer pair N8-1 and N8-2 respectively, it has the sequence as shown in SEQIDNo.1 to SEQIDNo.4 respectively.
2. for the identification of a primer pair for H10 subtype avian influenza virus, comprise Auele Specific Primer H10-1 and H10-2, they have the base sequence of sequence table SEQ IDNo.1 to SEQIDNo.2 respectively.
3. for the identification of a primer pair for N8 subtype avian influenza virus, comprise Auele Specific Primer N8-1 and N8-2, they have the base sequence of sequence table SEQ IDNo.3 to SEQIDNo.4 respectively.
4. a H10N8 subtype avian influenza virus duplex RT-PCR detection kit, comprise Reverse Transcription, PCR reagent, DEPC water and the ultrapure water without RNA enzyme, it is characterized in that: containing 2 pairs of Auele Specific Primers in described PCR reagent, be primer pair H10-1 and H10-2, primer pair N8-1 and N8-2 respectively, it has the sequence as shown in SEQIDNo.1 to SEQIDNo.4 respectively.
5. H10N8 subtype avian influenza virus duplex RT-PCR detection kit according to claim 4, is characterized in that: described Auele Specific Primer H10-1 and H10-2, N8-1 and the N8-2 final concentration in described PCR reagent is 0.1-0.8 μM.
6. H10N8 subtype avian influenza virus duplex RT-PCR detection kit according to claim 4, it is characterized in that: described Reverse Transcription is pipe 16 μ L often, wherein 5 × ReverseTranscriptaseBuffer10 μ L, 50pmol9merRandomPrimer2 μ L, 10mMdNTPMixture2 μ L, 40URibonucleaseInhibitor1 μ L, 5U/ μ LAMVReverseTranscriptase1 μ L.
7. H10N8 subtype avian influenza virus duplex RT-PCR detection kit according to claim 4, it is characterized in that: the volume of described PCR reagent is that 26.2 μ L/ manage, wherein comprise 2 × EasyTaqTMPCRSuperMix25 μ L, 50 μMs of each 0.2 μ L of H10-1 and H10-2 primer, 50 μMs of each 0.4 μ L of N8-1 and N8-2 primer.
8. the primer sets for the identification of H10N8 subtype avian influenza virus according to claim 1, the arbitrary described primer pair of claim 2-3 or the arbitrary described H10N8 subtype avian influenza virus duplex RT-PCR detection kit of claim 4-7 are differentiating the application in H10N8 subtype avian influenza virus.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106554413A (en) * | 2015-09-30 | 2017-04-05 | 复旦大学 | Full human monoclonal antibody and application for H10N8 subtype avian influenza virus |
| CN106702028A (en) * | 2017-02-27 | 2017-05-24 | 解码（上海）生物医药科技有限公司 | Real-time fluorescent PCR detection kit for H10N7 avian influenza virus and detection method of real-time fluorescent PCR detection kit |
| CN107858454A (en) * | 2017-12-06 | 2018-03-30 | 防城港市动物疫病预防控制中心 | H1N6 subtype avian influenza virus double RT PCR detection primers group, kit and its application |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103194443A (en) * | 2013-04-01 | 2013-07-10 | 青岛农业大学 | Multiple reverse transcription amplification method of three genes of H9N2 avian influenza virus |
-
2015
- 2015-12-25 CN CN201510988413.XA patent/CN105368988A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103194443A (en) * | 2013-04-01 | 2013-07-10 | 青岛农业大学 | Multiple reverse transcription amplification method of three genes of H9N2 avian influenza virus |
Non-Patent Citations (4)
| Title |
|---|
| 傅伟杰等: "江西省3例H10N8禽流感病毒感染者回归性分析", 《中华流行病学杂志》 * |
| 杨成民等: "《基础输血》", 31 October 2001, 北京：中国科学技术出版社 * |
| 武惠娟等: "《临床科研常用分子生物学技术》", 31 July 2015 * |
| 薄洪等: "H10N8禽流感病毒REAL TIME RT-PCR检测方法的建立", 《中华实验和临床病毒学杂志》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106554413A (en) * | 2015-09-30 | 2017-04-05 | 复旦大学 | Full human monoclonal antibody and application for H10N8 subtype avian influenza virus |
| CN106554413B (en) * | 2015-09-30 | 2020-07-14 | 复旦大学 | Fully human monoclonal antibody aiming at H10N8 subtype avian influenza virus and application |
| CN106702028A (en) * | 2017-02-27 | 2017-05-24 | 解码（上海）生物医药科技有限公司 | Real-time fluorescent PCR detection kit for H10N7 avian influenza virus and detection method of real-time fluorescent PCR detection kit |
| CN107858454A (en) * | 2017-12-06 | 2018-03-30 | 防城港市动物疫病预防控制中心 | H1N6 subtype avian influenza virus double RT PCR detection primers group, kit and its application |
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Application publication date: 20160302 |