Multiplex nested PCR for Brazilian Alphavirus diagnosis

被引：28

作者：

Bronzoni, RVM

Moreli, ML

Cruz, ACR

Figueiredo, LTM ^{[1
]}

机构：

[1] Univ Sao Paulo, Sch Med Ribeirao Preto, Virus Res Unit, BR-14049900 Ribeirao Preto, SP, Brazil

[2] Evandro Chagas Inst, Arbovirus Lab, Belem, Para, Brazil

来源：

TRANSACTIONS OF THE ROYAL SOCIETY OF TROPICAL MEDICINE AND HYGIENE | 2004年 / 98卷 / 08期

基金：

巴西圣保罗研究基金会;

关键词：

Alphavirus; reverse transcriptase; PCR; multiplex nested PCR; Brazil;

D O I：

10.1016/j.trstmh.2003.09.002

中图分类号：

R1 [预防医学、卫生学];

学科分类号：

1004 ; 120402 ;

摘要：

A reverse-transcriptase PCR (RT-PCR) and a multiplex nested PCR were developed for the rapid detection and identification of 14 Brazilian alphaviruses. Using Alphavirus genus-specific primers in a RT-PCR, we obtained amplified products of 434 bp. Species-specific primers were selected and simultaneously tested in a multiplex nested PCR. The nested PCR increased the test sensitivity 1000-fold and was capable of identifying Brazilian Alphavirus showing the expected bands with diagnostic sizes for Venezuelan (400 bp), Eastern (124 bp), and Western (208 bp) equine encephalitis, Aura (86 bp), and Mayaro (270 bp) viruses. This strategy for diagnosis is fast, sensitive, specific and it can be used as a reliable alternative for routine Brazilian Alphavirus diagnosis. (C) 2004 Royal Society of Tropical Medicine and Hygiene. Published by Elsevier Ltd. ALI rights reserved.

引用

页码：456 / 461

页数：6

共 16 条

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