牦牛星状病毒的RT-PCR检测方法的建立和变异分析

doi:10.11843/j.issn.0366-6964.2016.06.027

Abstract

Abstract:

Recently，our laboratory identified a novel astrovirus from fecal samples of diarrheic yak，the aim of this study was to develop an RT-PCR assay for detecting the novel yak astrovirus (yak AstV).Firstly，one pair primers were designed to amplify ORF2 gene fragments (630 bp in length) of yak AstV.By sequence analysis，one pair primers of RT-PCR assay were then designed for detecting yak AstV.We obtained 13 ORF2 gene fragments from 20 fecal samples of diarrheic yak，which shared a homology of 99.3%-100% within fragments，but only a homology of 59.5％-80.8％ with bovine astrovirus (BAstV).The RT-PCR assay developed in this study was specific to yak astrovirus，no amplifying for BAstV and other pathogens tested in this study.The detection limit of viral nucleic acid of the assay was 57.3 fg•μL^－1.The assay was significantly better in detecting yak AstV compared to the other two reported assays for detecting bovine astrovirus.The detecting results of clinical samples by this assay showed that the positive rates of yak AstV were 55.5% (76/137) in diarrheic fecal samples and 7.7％ (2/26) in healthy fecal samples.Compared to healthy samples，yak AstV was significantly more prevalent in diarrheic fecal samples (P<0.01).In conclusion，ORF2 gene sequences of yak AstV cloned in this study varied a lot compared with those of BAstV，but were highly conserved within yak AstV species.Thus，the RT-PCR for detecting yak AstV based on the ORF2 sequences of yak AstV greatly improved the detection rate of yak AstV，providing a useful tool for detection of this novel virus and epidemiological investigation.Meanwhile，the results of this study indicated that yak AstV might be associated with diarrhea in yak calves.

CLC Number:

S852.653

CHEN Xi，TANG Cheng，ZHANG Bin，SONG Zhi-gang，ZHOU Fang，YUE Hua. Development of an RT-PCR Assay and Variation Analysis for Yak Astrovirus[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2016, 47(6): 1287-1292.

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Development of an RT-PCR Assay and Variation Analysis for Yak Astrovirus

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