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通过逆转录聚合酶链反应(RT-PCR)快速检测实验室感染的埃及伊蚊中的基孔肯雅病毒。

Rapid detection of chikungunya virus in laboratory infected Aedes aegypti by Reverse-Transcriptase- Polymerase Chain Reaction (RT-PCR).

作者信息

Rohani A, Yulfi H, Zamree I, Lee H L

机构信息

Medical Entomology Unit, Infectious Disease Research Centre, Institute for Medical Research, Kuala Lumpur, Malaysia.

出版信息

Trop Biomed. 2005 Dec;22(2):149-54.

Abstract

A study of chikungunya virus was carried out to establish Reverse Transcriptase- Polymerase Chain Reaction (RT-PCR) as a rapid detection technique of the virus. The susceptibility of lab-colonized Aedes aegypti to chikungunya virus was also determined. Artificial membrane feeding technique was used to orally feed the mosquitoes with a human isolate of chikungunya virus. A total of 100 fully engorged female Ae. aegypti were obtained and maintained for 7 days. Seventy of them survived and then pooled at 10 individuals per pool. Total RNA was extracted from the samples and RT-PCR amplifications were carried out. Five out of 7 pools showed positive PCR band at 350-bp, indicating Ae. aegypti is a potential vector of chikungunya virus. The minimum infection rate (MIR) was 71% within these laboratory colonies. RT-PCR is a sensitive technique that is useful in detecting infected mosquitoes in epidemic areas. This technique can de used as a rapid detection method and provide an early virologic surveillance systems of chikungunya virus infected mosquitoes.

摘要

开展了一项关于基孔肯雅病毒的研究,以确立逆转录聚合酶链反应(RT-PCR)作为该病毒的快速检测技术。还测定了实验室饲养的埃及伊蚊对基孔肯雅病毒的易感性。采用人工膜饲技术,用一株基孔肯雅病毒的人类分离株经口喂养蚊子。总共获得了100只饱血的雌性埃及伊蚊,并饲养7天。其中70只存活下来,然后每10只分为一组。从样本中提取总RNA并进行RT-PCR扩增。7组中有5组在350bp处显示出阳性PCR条带,表明埃及伊蚊是基孔肯雅病毒的潜在传播媒介。在这些实验室群体中,最低感染率(MIR)为71%。RT-PCR是一种灵敏的技术,可用于检测流行地区受感染的蚊子。该技术可作为一种快速检测方法,并为基孔肯雅病毒感染蚊子提供早期病毒学监测系统。

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