摘要
本研究以鸡传染性支气管炎病毒(IBV)标准毒株M41为原料,通过SPF鸡胚尿囊腔繁殖、外源病毒检测、灭活、冻干和分装,添加适量的稳定剂和保护剂,最终成功制备鸡传染性支气管炎病毒核酸检测专用标准物质。采用荧光定量RT-PCR方法,由9家权威实验室进行合作定值,确定标准物质的定性检测结果和参考值。结果显示:所研制的标准物质纯净,均匀性和稳定性良好,定性准确,易保存,可常规运输,安全,无生物传染风险;标准物质在-20℃、4℃、25℃、37℃环境下可分别稳定保存12个月以上、60 d、7 d、3 d;合作定值验证和临床试用结果均为鸡传染性支气管炎病毒阳性。结果表明,所制备的标准物质可用于鸡传染性支气管炎病毒核酸检测相关实验室的质量检定和控制、仪器测试和校准、检测方法评价等,有利于鸡传染性支气管炎的诊断和防控。
In order to prepare qualitative reference material(RM)for detection of nucleic acid of infectious bronchitis virus(IBV),the classical strain M41 of IBV was used as the raw material,and a serial of experiments were performed,including inoculation of IBV into SPF chicken embryos,detection of exogenous viruses,inactivation,lyophilization,allocation,etc.Real time RT-PCR was used by our laboratory and 9 cooperating laboratories to qualitatively detect the RM.The results showed that the prepared RM was pure,stable,homogeneous,easy to preserve and transport,safe and no risk of biological infection.The results of stability evaluation of the RM indicated that it was stable when stored at-20℃,4℃,25℃and 37℃for 12 months,60 days,7 days and 3 days,respectively.Cooperating detection showed that all RMs examined were positive for IBV.The prepared RM is suitable to be used for quality control,comparison and evaluation of instruments and methods in laboratories relevant to nucleic acid detection of IBV,and would be beneficial to the diagnosis and control of IB.
作者
黄袁慧
王乃迪
陈玉红
高晓艺
刘洋
顾小雪
杨林
辛盛鹏
王传彬
刘玉良
HUANG Yuan-hui;WANG Nai-di;CHEN Yu-hong;GAO Xiao-yi;LIU Yang;GU Xiao-xue;YANG Lin;XIN Sheng-peng;WANG Chuan-bin;LIU Yu-liang(China Animal Disease Control Center,Beijing 102618,China;College of Animal Science and Technology,Guangxi University,Nanning 530005,China;School of Life Sciences and Food Engineering,Heibei University of Engineering,Handan 056038,China)
出处
《中国兽医杂志》
CAS
北大核心
2021年第5期60-64,69,I0006,共7页
Chinese Journal of Veterinary Medicine
基金
科技部科技基础性工作专项(2013FY113300-5)
国家重点研发计划项目(2016YFD0501609)。
关键词
鸡传染性支气管炎病毒
标准物质
核酸
检测
avian infectious bronchitis virus
reference material
nucleic acid
detection
