[一种用于检测腮腺炎病毒基因的SYBR Green实时逆转录聚合酶链反应的开发]

[Development of a SYBR green real-time RT-PCR for detection of mumps virus gene].

作者信息

Cui Ai-Li, Zhu Zhen, Wang Chang-Yin

机构信息

State Key Laboratory for Molecular Virology and Genetic, Engineering, Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100050, China.

出版信息

Zhongguo Yi Miao He Mian Yi. 2009 Feb;15(1):14-8.

PMID:20077669

Abstract

OBJECTIVE

A SYBR Green real-time RT-PCR assay was developed to detect mumps virus gene rapidly.

METHODS

The primers were selected based on SH gene of mumps virus, the assay was optimized in reactive system and condition to improve the sensitivity and specificity. The SH gene of 5 mumps virus isolates were detected and genotyped using this assay.

RESULTS

A SYBR Green real-time RT-PCR assay had good sensitivity and specificity. There was no cross reaction with other respiratory virus. The detection limit of the assay was 10 TCID50. The sensitivity of the two assays, RT-PCR and real-time RT-PCR, was not different. Compared with traditional RT-PCR, real-time RT-PCR saved half detection time. The products of SYBR Green real-time RT-PCR were directly sequenced and genotyped.

CONCLUSIONS

SYBR Green real-time RT-PCR was a rapid, specific and sensitive method for the detection of mumps virus and genotyping.

摘要

目的

建立一种SYBR Green实时逆转录聚合酶链反应（RT-PCR）检测方法，用于快速检测腮腺炎病毒基因。

方法

根据腮腺炎病毒的SH基因选择引物，对反应体系和条件进行优化以提高敏感性和特异性。使用该检测方法对5株腮腺炎病毒分离株的SH基因进行检测和基因分型。

结果

SYBR Green实时RT-PCR检测方法具有良好的敏感性和特异性。与其他呼吸道病毒无交叉反应。该检测方法的检测限为10个半数组织培养感染剂量（TCID50）。RT-PCR和实时RT-PCR这两种检测方法的敏感性无差异。与传统RT-PCR相比，实时RT-PCR节省了一半的检测时间。SYBR Green实时RT-PCR的产物直接进行测序和基因分型。