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用于快速检测马尔堡病毒和埃博拉病毒的多重实时聚合酶链反应方法

[Multiplex real-time PCR method for rapid detection of Marburg virus and Ebola virus].

作者信息

Yang Yu, Bai Lin, Hu Kong-Xin, Yang Zhi-Hong, Hu Jian-Ping, Wang Jing

机构信息

Chinese Academy of Inspection and Quarantine, Beijing 100123, China.+

出版信息

Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi. 2012 Aug;26(4):313-5.

Abstract

OBJECTIVE

Marburg virus and Ebola virus are acute infections with high case fatality rates. A rapid, sensitive detection method was established to detect Marburg virus and Ebola virus by multiplex real-time fluorescence quantitative PCR.

METHODS

Designing primers and Taqman probes from highly conserved sequences of Marburg virus and Ebola virus through whole genome sequences alignment, Taqman probes labeled by FAM and Texas Red, the sensitivity of the multiplex real-time quantitative PCR assay was optimized by evaluating the different concentrations of primers and Probes.

RESULTS

We have developed a real-time PCR method with the sensitivity of 30.5 copies/microl for Marburg virus positive plasmid and 28.6 copies/microl for Ebola virus positive plasmids, Japanese encephalitis virus, Yellow fever virus, Dengue virus were using to examine the specificity.

CONCLUSIONS

The Multiplex real-time PCR assays provide a sensitive, reliable and efficient method to detect Marburg virus and Ebola virus simultaneously.

摘要

目的

马尔堡病毒和埃博拉病毒是病死率很高的急性感染性疾病。通过多重实时荧光定量PCR建立一种快速、灵敏的检测方法来检测马尔堡病毒和埃博拉病毒。

方法

通过全基因组序列比对,从马尔堡病毒和埃博拉病毒的高度保守序列设计引物和Taqman探针,用FAM和Texas Red标记Taqman探针,通过评估不同浓度的引物和探针优化多重实时定量PCR检测方法的灵敏度。

结果

我们开发了一种实时PCR方法,对马尔堡病毒阳性质粒的灵敏度为30.5拷贝/微升,对埃博拉病毒阳性质粒的灵敏度为28.6拷贝/微升,用日本脑炎病毒、黄热病毒、登革病毒检测其特异性。

结论

多重实时PCR检测方法为同时检测马尔堡病毒和埃博拉病毒提供了一种灵敏、可靠且高效的方法。

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