Hendra病毒的实时PCR检测方法的建立

Detection of Hendra virus by real-time PCR assay

目的:建立针对Hendra病毒的敏感、特异和快速的实时PCR法,为Hendra病的早期诊断提供依据。方法:采用常规PCR法和实时PCR法分别对构建的Hendra病毒全长G基因的重组质粒DNA进行扩增,并比较两种方法的敏感性。结果与结论:应用常规PCR方法可检测到100fg/μl的含Hendra病毒特异序列的重组质粒DNA,而实时PCR法则可检测到0.1fg/μl的DNA,后者的敏感性比常规PCR法的提高了1000倍,该方法不但操作上更加简便、快速,还可避免交叉污染,适于对Hendra病毒特异序列的检测。

Objective:To develop a sensitive, specific and rapid real-time PCR assay for early diagnosis of Hendra disease. Methods: Recombinant plasmid DNA of Hendra virus G gene were amplified by conventional PCR and real-time PCR assays, and sensitivity of the two methods was compared. Results and Conclusion: 100 fg/μl of recombinant plasmid DNA of Hendra virus specific sequences were detected by conventional PCR assay, 0.1 fg/μl of DNA was detected by real-time PCR assay,the sensitivity of the latter was 1000-fold greater than conventional one.This assay was not only more simple,time-saving in operation, but also could avoid cross-contamination, thus could be used for detection of specific sequences of Hendra virus.