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应用 RT-PCR 技术从快速层析检测试纸条中检测牛瘟病毒 RNA。

The detection of rinderpest virus RNA extracted from a rapid chromatographic strip-test by RT-PCR.

机构信息

Institute for Animal Health Pirbright Laboratory, Ash Road, Pirbright, Surrey GU 21 3 LL, United Kingdom.

出版信息

J Virol Methods. 2011 May;173(2):394-8. doi: 10.1016/j.jviromet.2011.02.020. Epub 2011 Feb 23.

Abstract

The Global Rinderpest Eradication Program (GREP) aimed to eradicate rinderpest by 2010 and it is widely believed to have been successful. An integral part of the program was the submission of samples from suspect rinderpest positive animals to a local Reference Laboratory for final confirmation. Confirmation of rinderpest in field samples is often hampered because of poor quality of the sample upon receipt. As part of GREP a rapid diagnostic strip test for the detection of rinderpest virus (RPV) in the field was developed allowing a rapid response to suspect outbreaks. The feasibility of extracting viral RNA from the used rapid diagnostic rinderpest devices for final confirmation in the laboratory is described. Viral material contained within used rinderpest devices was stable enough after storage for one week at 21°C to extract RNA from five different RPV strains and amplify it by reverse transcriptase polymerase chain reaction (RT-PCR). Temperature did not affect adversely the extraction and amplification of the viral RNA but humidity impaired RNA extraction and amplification. Used rinderpest devices from field diagnosed rinderpest-positive animals could represent an ideal additional sample for submission to the Reference Laboratories for confirmation of preliminary diagnosis in the field.

摘要

全球牛瘟根除计划(GREP)旨在到 2010 年根除牛瘟,人们普遍认为该计划已经取得成功。该计划的一个组成部分是将来自疑似牛瘟阳性动物的样本提交给当地参考实验室进行最终确认。由于收到的样本质量较差,因此在现场确认牛瘟时经常会遇到困难。作为 GREP 的一部分,开发了一种用于现场检测牛瘟病毒(RPV)的快速诊断带测试,以便对疑似暴发做出快速反应。本文描述了从已使用的快速诊断牛瘟设备中提取病毒 RNA 以在实验室进行最终确认的可行性。在 21°C 下储存一周后,储存在已使用的牛瘟设备内的病毒材料足够稳定,可从五种不同的 RPV 株中提取 RNA 并通过逆转录聚合酶链反应(RT-PCR)进行扩增。温度不会对病毒 RNA 的提取和扩增产生不利影响,但湿度会影响 RNA 的提取和扩增。来自现场诊断为牛瘟阳性动物的已使用牛瘟设备可以作为提交给参考实验室的理想补充样本,以确认现场初步诊断。

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