Novel goose arbovirus detection primer group and kit

A technique for detection of astroviruses and primers, applied in the field of real-time fluorescent quantitative PCR detection primers and its kits, can solve the problems of sample cross-contamination, time-consuming and labor-intensive, low reproduction titer, etc., and achieve strong specificity and good repeatability , detect the effect of fast

Pending Publication Date: 2020-05-01
SOUTH CHINA AGRI UNIV
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Problems solved by technology

Regarding the diagnosis of the virus, the traditional virus isolation and identification is time-consuming and laborious, and is not suitable for clinical use. Moreover, the virus has no hemagglutination, does not grow on chicken embryos, duck embryos, and the reproductive titer on g...

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  • Novel goose arbovirus detection primer group and kit
  • Novel goose arbovirus detection primer group and kit
  • Novel goose arbovirus detection primer group and kit

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Embodiment Construction

[0030] 1. Related test pathogens

[0031] The pathogenic goose Astrovirus, goose-derived Escherichia coli, goose-derived Riemerella anatipestifer, goose parvovirus, goose circovirus and H9N2 subtype avian influenza were identified and approved by the Poultry Disease Teaching and Research Group of the School of Veterinary Medicine, South China Agricultural University. save.

[0032] 2. Primer design

[0033] According to the gene sequence fragment encoded by the goose astrovirus (goose astrovirus GoAstV QY / 126 strain) polymerase 2 protein (ORF2 gene) previously identified by our research group, the specific primers for goose astrovirus were designed as follows:

[0034] Upstream primer GoAstV-F1: 5'-CCCTTGTAAATACCCAGGATG-3',

[0035] Downstream primer GoAstV-R1: 5'-CTCCAAGTGCACTCGCTACG-3',

[0036] The target fragment size is 174bp;

[0037] Primer BLAST analysis was performed in the NCBI database to verify the specificity of the designed primers.

[0038] The above primers ...

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Abstract

The invention discloses real-time fluorescence quantification PCR (polymerase chain reaction) detection primers for a novel goose arbovirus which causes gout of goslings, and a kit of the primers. Thekit comprises an upstream primer GoAstV-F1 and a downstream primer GoAstV-R1. The kit disclosed by the invention is capable of detecting the novel goose arbovirus which causes gout of the goslings, has the advantages of being good in specificity, good in repeatability and accurate in quantification, is good in sensitivity, is capable of detecting 6.8 copies at least, has a very good linear relationship within a 6.88-6.88*10<9> copy range of a standard product, has a detection range up to 10 orders of magnitudes, is capable of rapidly detecting the novel goose arbovirus GoAstV which causes gout of the goslings, and improves the detection efficiency.

Description

technical field [0001] The invention belongs to the field of gene detection, and in particular relates to a real-time fluorescent quantitative PCR detection primer and a kit thereof for a novel goose astrovirus. Background technique [0002] Astrovirus (Arbovirus) is a single-stranded positive-sense RNA virus belonging to the family Astroviridae. The genome length is 6.1-7.9kb, and the genome includes 5'UTR, ORF1a, ORF1b and ORF2 three open reading frames and 3'URT , polyadenylic acid (PolyA) tail. The virus infection has a certain prevalence, and can cause enteritis and diarrhea in humans or animals, accompanied by vomiting and diarrhea. Astroviridae can be divided into mammalian genera and avian genera according to different hosts, among which mammals are divided into human astrovirus, feline astrovirus, porcine astrovirus, sheep astrovirus and mink astrovirus according to different hosts. Astroviruses; the genus Astrovirus includes duck Astrovirus (DAstV), turkey Astrov...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/686C12N15/11
CPCC12Q1/701C12Q1/686C12Q2561/113C12Q2563/107C12Q2545/114
Inventor 罗开健施建鑫冯赛祥王晓冰林桢毓李伟业陈爱华
Owner SOUTH CHINA AGRI UNIV
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