摘要
目的建立针对西尼罗病毒E基因的一步法Real-time RT-PCR检测方法,用于西尼罗病毒感染样本的快速准确检测和定量。方法针对西尼罗病毒的保守基因E设计引物和探针,建立一步法Real-time RT-PCR反应方法并分析敏感性和特异性。结果本研究建立的一步法Real-time RT-PCR方法可以特异性检测出西尼罗病毒,不与日本脑炎病毒及其他病毒产生交叉反应。该方法的最低检出浓度为3.6×102copies/μl,标准曲线的线性范围为3.6×102～3.6×107copies/μl。结论本研究建立的一步法real-time RT-PCR方法敏感性和特异性较高,且不易出现污染引起的假阳性结果,适合用于西尼罗病毒感染样本的检测。
One step real-time RT-PCR（Taqman） assays of West Nile virus（WNV） envelope protein gene was developed so that WNV RNA in field specimens or laboratory material can be characterized rapidly and specifically. A pair of specific oligonucleotide primers was designed. The linearity of the standard curve allowed quantification of 10^2 to 107 RNA transcripts/ul. and sensitivity of the test was close to 3.6 ×10^2 transcripts/ul. This assays were high specific without cross-reaction to Japanese encephalitis virus or other virus. This standardized technique is sensitive and reliable and allows rapid detection and quantiration of West Nile virus RNA in both field and experimental materials used for the surveillance and specific diagnosis of West Nile virus.
出处
《中国人兽共患病学报》
CAS
CSCD
北大核心
2008年第12期1107-1110,共4页
Chinese Journal of Zoonoses
基金
"十一五"国家高科技发展计划(863计划)(No.2006AA02Z196)资助
