[应用Taqman MGB实时荧光定量PCR技术快速检测登革病毒的研究]

[Study on the rapid detection of Dengue viruses by Taqman MGB real-time fluorescent PCR].

作者信息

Liu Jian-jun, Yang Fan, He Jian-fan, Chen Jia-min, He Ya-qing, Yang Hong

机构信息

Shenzhen Center for Disease Control and Prevention, Shenzhen 518020, China.

出版信息

Wei Sheng Yan Jiu. 2006 Nov;35(6):736-8.

PMID:17290754

Abstract

OBJECTIVE

To develop the Taqman MGB real-time fluorescent PCR assay for rapid detection of Dengue viruses.

METHODS

Using Taqman MGB technique, a pair of universal primers and Taqman MGB probe were designed according to a highly reserved sequence of the 3'-noncoding region of dengue viruses type 1-4. Dengue virus strains were used as standard and Japanese encephalitis virus strains were used as control, the real-time PCR assay for specific and sensitive detection of the dengue viruses was established. While 8 serum specimens of ELISA positive were detected by the RT-PCR and fluorescent PCR.

RESULTS

The sensitivity of real-time PCR was 0.17pg/microl (cDNA)or 10(-5)TCID50. There was no cross-reaction with Japanese encephalitis virus. Of 8 specimens, 2 were positive by RT-PCR and 5 were positive by real-time PCR. The test could be completed in 4 hours.

CONCLUSION

The Taqman MGB real-time PCR assay was fast, sensitive and specific. It could be applied to the quick early diagnosis of dengue viruses.

摘要

目的

建立用于快速检测登革病毒的Taqman MGB实时荧光定量PCR方法。

方法

运用Taqman MGB技术，依据登革病毒1 - 4型3' - 非编码区高度保守序列设计一对通用引物及Taqman MGB探针。以登革病毒株为标准品，日本脑炎病毒株为对照，建立特异、灵敏检测登革病毒的实时荧光定量PCR方法。同时应用该方法对8份ELISA阳性血清标本进行RT - PCR及荧光定量PCR检测。