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Method and special reagent kit for detecting five zoonosis virus
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A kit and virus technology, which is applied in the field of detecting five kinds of zoonotic viruses, can solve the problems of manpower and time-consuming, complicated operation process, etc., and achieve the effects of saving time, high detection efficiency and reliable detection results
Inactive Publication Date: 2011-04-27
MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI
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However, the RT-PCR method can only detect one pathogen at a time. When it is used for the detection of multiple infections or unknown pathogens, multiple experimental reactions are required, and the operation process is complicated, labor-intensive and time-consuming.
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Embodiment 1
[0031] The design of the primer sequence of embodiment 1, multiplex RT-PCR
[0032] According to the genome sequences of FMDV, VSV, RVFV, RV and HEV provided by GenBank, the Bioedit analysis software was used for multiple sequence alignment analysis, and highly conserved segments were selected, respectively, the non-structural protein 3ABC region of FMDV, the N Protein region, M protein region of RVFV, structural protein ORF2 region of HEV, 5 pairs of specific primers were designed using Primer5.0 and AnnHyb multiple primer design software. The primer sequences and the lengths of the amplified gene products are listed in Table 1.
[0033] Table 1. Multiplex RT-PCR primer design
[0034]
Embodiment 2
[0035] Embodiment 2, detect virus with multiple RT-PCR primers
[0036] Foot and mouth disease virus (Foot and mouth disease virus, FMDV) was purchased from Harbin Veken Biotechnology Development Company;
[0037] Vesicular stomatitis virus (VSV) was purchased from Harbin Veken Biotechnology Development Company;
[0038] Rift valley fever virus (Rift valley fever virus, RVFV) recombinant plasmid (Wang Qinghua, Wang Xijun, Hu Sen, Bu Zhigao recombinant baculovirus expression of Rift valley fever capsule membrane protein [J]. Chinese Journal of Preventive Veterinary Medicine, 2006, 1( 28): 22-25) (Institute of Microbial Epidemiology, Academy of Military Medical Sciences, Chinese People's Liberation Army);
[0039] Rabies virus (RV) (Liu Zhongyu; Yuan Huijun; Zhu Wuyang; Jiang Tao; Chen Shuiping; Yu Man; Qin Chengfeng; Qin Ede; construction and identification of rabies virus vector, Chinese Journal of Zoonoses 2008 11 2008 , 24(11)) (Institute of Microbial Epidemiology, Academy...
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Abstract
The invention discloses a method for diagnosing five varieties of disease virus infected by both human beings and animals and a special kit thereof. The kit comprises a primer set A used for diagnosing foot and mouth disease virus: the DNA indicated by the sequence 1 in the sequence table, and the DNA indicated by the sequence 2 in the sequence table. The method of using the kit for diagnosing viruses has the advantages of high specificity and reliable results. When using a plurality of varieties of primers for diagnosing simultaneously, the diagnosis of the plurality of varieties of viruses in an identical PCR reaction can be achieved, the diagnosis efficiency is high, time and reagent are saved greatly and cost is reduced; therefore, the method has convenient, rapid and sensitive advantages, has significance for enhancing the monitoring of disease virus infected by both human beings and animals and for preventing the propagation of the disease virus infected by both human beings andanimals.
Description
technical field [0001] The invention relates to a method and a special kit for detecting five zoonosis viruses. Background technique [0002] In recent years, there has been a resurgence of zoonotic diseases around the world. Rift valley fever virus (RVFV), hepatitis E virus (Rabies virus, HEV), rabies virus (Rabies virus, RV), vesicular stomatitis virus (VSV) and Foot and mouth disease virus (FMDV) is an important pathogen of zoonotic infectious diseases. It has a wide range of epidemics, seriously endangers the development of animal husbandry, and also seriously threatens public health security. It is recognized by the World Organization for Animal Health (OIE) Classified as a class A disease. At present, the commonly used methods for detecting the above five viruses mainly include: animal inoculation, virus isolation and culture, enzyme-linked immunoassay (ELISA), serum neutralization test and other methods. In recent years, RT-PCR detection methods for the above virus...
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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/70C12Q1/68C12R1/93
Inventor 杨银辉孙庆歌吴东来康晓平李永强刘洪祝庆余
Owner MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI
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