逆转录聚合酶链反应-酶联免疫吸附测定法作为诊断低致病性禽流感的一种工具。

RT-PCR-ELISA as a tool for diagnosis of low-pathogenicity avian influenza.

作者信息

Dybkaer Karen, Munch Mette, Handberg Kurt J, Jørgensen Poul H

机构信息

Danish Veterinary Institute, Hangevej 2, DK-8200 Aarhus N, Denmark.

出版信息

Avian Dis. 2003;47(3 Suppl):1075-8. doi: 10.1637/0005-2086-47.s3.1075.

DOI:10.1637/0005-2086-47.s3.1075

PMID:14575114

Abstract

A one-tube reverse transcriptase/polymerase chain reaction coupled with an enzyme-linked immunosorbent assay (RT-PCR-ELISA) was developed for the rapid detection of avian influenza virus (AIV) in clinical specimens. A total of 419 swab pools were analyzed from chickens experimentally infected with low-pathogenicity AIV, from wild aquatic birds, and from domestic ducks. The AIV was detected in 32 swab pools by RT-PCR-ELISA compared to 23 by virus isolation (VI) in embryonated specific pathogen free (SPF) chicken eggs. Thus, 39% more specimens were positive by RT-PCR-ELISA than by VI. Two of the twenty-three VI-positive specimens were negative when tested by RT-PCR-ELISA. The diagnostic sensitivity and specificity of the RT-PCR-ELISA was 91% and 97%, respectively, using VI in SPF eggs as the gold reference standard.

摘要

开发了一种单管逆转录酶/聚合酶链反应与酶联免疫吸附测定相结合的方法（RT-PCR-ELISA），用于快速检测临床样本中的禽流感病毒（AIV）。对419份拭子样本池进行了分析，这些样本来自实验感染低致病性AIV的鸡、野生水鸟和家鸭。通过RT-PCR-ELISA在32份拭子样本池中检测到AIV，相比之下，在无特定病原体（SPF）的鸡胚中通过病毒分离（VI）检测到23份。因此，RT-PCR-ELISA检测出阳性的样本比VI多39%。23份VI阳性样本中有两份在通过RT-PCR-ELISA检测时呈阴性。以SPF鸡胚中的VI作为金标准参考，RT-PCR-ELISA的诊断敏感性和特异性分别为91%和97%。