Traul Donald L, Taus Naomi S, Lindsay Oaks J, O'Toole Donal, Rurangirwa Fred R, Baszler Timothy V, Li Hong
Animal Disease Research Unit, USDA-Agricultural Research Service, 3003 ADBF, Washington State University, Pullman, WA 99164, USA.
J Vet Diagn Invest. 2007 Jul;19(4):405-8. doi: 10.1177/104063870701900412.
Sheep-associated malignant catarrhal fever (SA-MCF), a frequently fatal disease primarily of certain ruminants, is caused by ovine herpesvirus 2 (OvHV-2). Molecular diagnosis of SA-MCF in affected animals has relied on detection of OvHV-2 DNA using a nested PCR, which has significant potential for amplicon contamination as a routine method in diagnostic laboratories. In this report, a nonnested and a previously developed real-time PCR were validated for detection of OvHV-2 DNA in samples from clinically affected animals. Three sets of blood or tissue samples were collected: 1) 97 samples from 97 naturally affected animals with evidence of clinical SA-MCF; 2) 200 samples from 8 animals with experimentally induced SA-MCF; and 3) 100 samples from 100 animals without any evidence of clinical SA-MCF. Among 97 positive samples defined by nested PCR from clinically affected animals, 95 (98%) were positive by nonnested PCR and 93 (96%) were positive by real-time PCR, respectively. One hundred percent of the samples from the animals with experimentally induced MCF were positive by real-time PCR, while 99% were positive by nonnested PCR. Neither nonnested PCR nor real-time PCR yielded a positive result on any of the 100 nested PCR-negative samples from animals without evidence of clinical MCF. The data confirmed that both nonnested and real-time PCR maintained high specificity and sensitivity for the detection of OvHV-2 DNA in clinical samples.
绵羊相关的恶性卡他热（SA-MCF）是一种主要发生于某些反刍动物的常见致命疾病，由绵羊疱疹病毒2型（OvHV-2）引起。对受感染动物进行SA-MCF的分子诊断依赖于使用巢式PCR检测OvHV-2 DNA，而作为诊断实验室的常规方法，该方法存在扩增子污染的重大风险。在本报告中，对一种非巢式PCR和一种先前开发的实时PCR进行了验证，用于检测临床受感染动物样本中的OvHV-2 DNA。收集了三组血液或组织样本：1）来自97只自然感染且有临床SA-MCF证据的动物的97份样本；2）来自8只经实验诱导感染SA-MCF的动物的200份样本；3）来自100只无任何临床SA-MCF证据的动物的100份样本。在巢式PCR定义的97份临床受感染动物的阳性样本中，分别有95份（98%）通过非巢式PCR呈阳性，93份（96%）通过实时PCR呈阳性。经实验诱导感染MCF的动物样本，100%通过实时PCR呈阳性，而99%通过非巢式PCR呈阳性。对于来自无临床MCF证据动物的100份巢式PCR阴性样本，非巢式PCR和实时PCR均未得出阳性结果。数据证实，非巢式PCR和实时PCR在检测临床样本中的OvHV-2 DNA时均保持了高特异性和高灵敏度。
