牛冠状病毒RT-PCR检测方法的建立及应用

doi:10.11843/j.issn.0366-6964.2018.10.026

Abstract

Abstract:

The aim of this study was to establish a more sensitive RT-PCR assay for detecting BCoV and to detect CoV from clinically ill yak in the northwest grasslands of Sichuan. The RT-PCR assay was established through designing primers targeted to Nsp7-Nsp9 fragment of BCoV polymerase gene and optimizing the reaction conditions and system, and the clinical samples were detected by the RT-PCR assay. Results revealed that the RT-PCR assay have good specificity and stability, and the detection limit of viral nucleic acid of the assays was 1×10^-2pg·μL^-1. Comparing to two RT-PCR assays targeted to the N gene of BCoV, the RT-PCR assay in this study has a remarkable detection rate for BCoV in clinical samples both of bovine and yak. In 125 diarrhea samples of yak in the northwest grasslands of Sichuan in 2016, the CoV detection rate was 71.20%, In 98 nasal cavity samples of yak in the northwest grasslands of Sichuan in 2016, the CoV detection rate was 72.45%. The RT-PCR assay for detecting BCoV established in this study has a good specificity and stability. BCoV is an important causative agent of yak diarrhea and respiratory disease syndrome in the northwest grasslands of Sichuan.

CLC Number:

S852.659.6

HE Qi-fu, GUO Zi-jing, LI Ran, ZHOU Jun, YUE Hua, ZHANG Bin, TANG Cheng. Establishment and Application of a RT-PCR Assay for Detecting Bovine Coronavirus[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(10): 2292-2298.

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Establishment and Application of a RT-PCR Assay for Detecting Bovine Coronavirus

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