Breslin J J, Smith L G, Barnes H J, Guy J S
Department of Microbiology, Pathology, and Parasitology, North Carolina State University, Raleigh 27606, USA.
Avian Dis. 2000 Jul-Sep;44(3):624-31.
A reverse transcriptase-polymerase chain reaction (RT-PCR) procedure and two monoclonal antibody (MAb)-based immunohistochemical procedures were developed for detection of turkey coronavirus (TCV) in tissues and intestinal contents/dropping samples. The RT-PCR, MAb-based fluorescent antibody (FA), and MAb-based immunoperoxidase (IP) procedures were compared with virus isolation (VI) for detection of TCV in experimentally infected turkeys. TCV was detected in experimentally infected turkeys as early as day 1 postexposure (PE) by each of the four detection procedures. TCV was detected as late as day 35 PE by FA or IP and days 42 and 49 PE by VI and RT-PCR, respectively. With VI as a reference, sensitivity and specificity of RT-PCR were 93% and 92%, respectively; specificity of both FA and IP was 96%, and sensitivities were 69% and 61%, respectively. Each of the examined procedures was highly specific, but the RT-PCR procedure was also highly sensitive. These findings demonstrate the utility of both immunohistochemistry and RT-PCR for detection of TCV. In addition, the findings indicate that RT-PCR is a highly sensitive and specific alternative to conventional diagnostic procedures.
已开发出一种逆转录酶-聚合酶链反应(RT-PCR)方法以及两种基于单克隆抗体(MAb)的免疫组织化学方法,用于检测组织和肠道内容物/粪便样本中的火鸡冠状病毒(TCV)。将RT-PCR、基于MAb的荧光抗体(FA)和基于MAb的免疫过氧化物酶(IP)方法与病毒分离(VI)进行比较,以检测实验感染火鸡中的TCV。通过这四种检测方法中的每一种,最早在暴露后第1天(PE)就检测到了实验感染火鸡中的TCV。通过FA或IP分别在最晚暴露后第35天检测到TCV,通过VI和RT-PCR分别在暴露后第42天和第49天检测到TCV。以VI作为参考,RT-PCR的敏感性和特异性分别为93%和92%;FA和IP的特异性均为96%,敏感性分别为69%和61%。所检测的每种方法都具有高度特异性,但RT-PCR方法也具有高度敏感性。这些发现证明了免疫组织化学和RT-PCR在检测TCV方面的实用性。此外,这些发现表明RT-PCR是传统诊断方法的一种高度敏感和特异的替代方法。