中国草食动物科学2025，Vol.45Issue(1)：44-49,6.DOI:10.3969/j.issn.2095-3887.2024.00.100
牛轮状病毒微滴式数字RT-PCR检测方法的建立及初步应用
Establishment and Application of Droplet Digital RT-PCR Detection Method for Bovine Rotavirus
摘要
Abstract
The purpose of this study was to establish a more sensitive detection method for bovine rotavirus(BRV),and to provide more accurate and sensitive technical support for the etiological investigation of BRV and the prevention and control of diseases in the livestock industry.According to the BRV VP6 genome sequence,specific primers and probes were designed and synthesized using Oligo7.56 software.The droplet digital RT-PCR method for BRV detection was established by optimizing the reaction conditions,while the specificity,sensitivity and repeatability tests were carried out,and the established method was applied to the detection of 14 clinical samples.The results showed that proposed method had a minimum detection limit of 1.83 copies/μL,which was reproducible and had no cross-reactivity with bo-vine infectious rhinotracheitis virus,bovine viral diarrhea virus,bovine coronavirus,bovine respiratory syncytial virus,and bovine parainfluenza virus type 3.Fourteen clinical samples were tested,and 4 samples were positive,with a posi-tive rate of 28.57%,and the coincidence rate between the test results and the RT-PCR test results was 100.00%.In su-mmary,the droplet digital RT-PCR method for the detection of BRV established in this study had good specificity,sen-sitivity and reproducibility,and could be applied to the detection of clinical samples,providing technical support for the diagnosis and epidemiological investigation of BRV.关键词
牛轮状病毒/VP6基因/微滴式数字RT-PCRKey words
bovine rotavirus/VP6 gene/droplet digital RT-PCR分类
畜牧业引用本文复制引用
谢世斌,郑辉,沙洲,房琳琳,尼博,张慧,董雅琴,魏荣,崔进..牛轮状病毒微滴式数字RT-PCR检测方法的建立及初步应用[J].中国草食动物科学,2025,45(1):44-49,6.基金项目
"十四五"国家重点研发计划项目(2021YFD1800300) （2021YFD1800300）
