Novel goose astrovirus and duck Tembusu virus multiplex fluorescence quantitative PCR detection primer probe set, kit and application
A technology for duck tembusu virus and astrovirus, which is applied in recombinant DNA technology, microbial determination/inspection, and microorganisms, etc., can solve the problems such as unreported multiple fluorescence quantitative PCR method, infection, and loss of goose breeding industry.
Active Publication Date: 2022-07-22
INST OF ANIMAL HUSBANDRY & VETERINARY MEDICINE JIANGXI ACAD OF AGRI SCI +1
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[0002] Since 2017, a number of goose farms in East China have had an outbreak of goose goose as the main clinical feature, mainly infecting goslings aged 4 to 16 days, resulting in a mortality rate of about 2% to 20%. Dead geese manifested as kidney disease. White swelling, pleural peritoneum, heart surface, and ureter have a large amount of urate deposition, which has caused great losses to the goose industry.
[0004] At present, there have been researches on the establishment of a fluorescent quantitative PCR method for Tembusu virus and the establishment of a fluorescent quantitative PCR detection method for the new duck parvovirus SYBRGreenΙ, but the multiplex fluorescent quantitative PCR method for the new goose Astrovirus and duck Tembusu has not been reported.
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[0044]In the present invention, primers and probes are successively designed for E gene of Tembusu virus (DTMUV), ORF1b gene, ORF1a gene and ORF2 gene of novel goose astrovirus (GoAstV), as shown in Table 1 and Table 2 below, a total of 9 pair primers and probes. The amplification effects of each pair of primers and probes in Table 2 are shown in order Figure 10-Figure 14 As shown, there is no amplified signal. However, only the primers and probe combinations designed for the E gene of Tembusu virus (DTMUV) and the ORF1b gene of the novel goose astrovirus (GoAstV) (Table 1) performed better. The primers and probes experimental scheme and experimental effects in Table 1 are described in detail below.
[0045] Table 1 Successful primer information
[0046]
[0047] Table 2 Failed primer information
[0048]
[0049] Fluorescence quantitative PCR experiments were performed according to the instructions of the kit Nanjing Novizan AceQ qPCR Probe Master Mix (Item No.: Q1...
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The invention belongs to the technical field of virus detection, and in particular relates to a novel goose astrovirus (GoAstV) and duck tembusu virus (DTMUV) multiplex fluorescence quantitative PCR detection primer probe set, kit and application. In this application, specific primers and probes were designed for the E gene of DTMUV and the ORF1b gene of GoAstV, respectively. After optimizing the reaction conditions, a fluorescent quantitative PCR method for simultaneous detection of DTMUV and GoAstV in the same system was established. This method has the advantages of specificity, sensitivity and high efficiency.
Description
technical field [0001] The invention belongs to the technical field of virus detection, and in particular relates to a novel goose astrovirus and duck Tembusu virus multiplex fluorescence quantitative PCR detection primer probe set, kit and application. Background technique [0002] Since 2017, many goose farms in East China have outbreaks with gosling as the main clinical feature, mainly infecting goslings aged 4 to 16 days, resulting in a mortality rate of about 2% to 20%. The dead geese manifested as kidney disease. White and swollen, pleural, peritoneal and cardiac surfaces, and a large number of urate deposits in the ureter, have caused great losses to the goose industry. After isolation and identification, it was found that the disease was caused by an astrovirus infection. This strain is very different from the previously reported avian astroviruses. Its whole genome homology is 46.5% to 62.0%, and the coding The homology of the amino acid sequence of the ORF2 gene o...
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IPC IPC(8): C12Q1/70C12Q1/6851C12N15/11C12R1/93
CPCC12Q1/701C12Q1/6851C12Q2600/16C12Q2531/113C12Q2537/143C12Q2561/101
Inventor 李海琴杨群张帆帆谭美芳康昭风傅秋玲曾艳兵季华员黄江南傅光华韦启鹏黄瑜谭佳吴诚诚方绍培
Owner INST OF ANIMAL HUSBANDRY & VETERINARY MEDICINE JIANGXI ACAD OF AGRI SCI