摘要
[目的]针对牛传染性鼻气管炎病毒建立快速、准确而敏感的分子生物学检测方法,为进出口牛及其遗传物质的IBR检疫把关提供新的技术手段。[方法]采用新型实时荧光PCR技术原理,自行设计荧光标记引物建立检测方法,对不同牛传染性鼻气管炎病毒样品进行敏感性、特异性等检测试验,并与常规PCR检测方法进行比较。[结果]该方法能特异检测IBR病毒,检测时间包括核酸样品制备仅需2h,敏感性比常规PCR检测方法提高达103-104。[结论]本方法适用于在牛及其遗传物质的进出口检疫中进行牛传染性鼻气管炎快速病原鉴定。
A rapid,accurate and sensitive method was studied to detect the infectious bovine rhinotracheitis virus(IBRV) for the quarantine of import & export cattle and genetic materials.A real-time PCR assay,based on LUX(Light Upon Extension) Fluorogenic Primer and LightCycle technology,was developed for detection of IBRV.It was shown specifically to detect viral DNA from different IBRV infected cell cultivates.The entire test could be finished within 2 hours,which included the time for nucleic acid extraction.Serial 10 fold dilution of IBRV DNA samples were detected by the assay and a published gel-based PCR method respectively.The result demonstrated a 103-4-fold increase of sensitivity of the real time assay than the gel-based method.The study indicates that the method reported here provides a reliable tool for rapid detection of IBRV in import & export cattle and genetic materials.
出处
《检验检疫科学》
2005年第z1期5-7,共3页
Inspection and Quarantine Science
关键词
牛传染性鼻气管炎
实时
荧光PCR
infectious bovine rhinotracheitis virus(IBRV)
real-time
fluorescence PCR
