基于聚合酶链反应的检测方法，用于检测来自受感染摩洛哥绵羊的眼拭子、鼻拭子和直肠拭子中的绵羊痘病毒。

PCR-based assay to detect sheeppox virus in ocular, nasal, and rectal swabs from infected Moroccan sheep.

作者信息

Zro K, Azelmat S, Bendouro Y, Kuhn J H, El Fahime E, Ennaji M M

机构信息

Laboratory of Virology, Microbiology and Quality/Ecotoxicology and Biodiversity, Hassan II University Mohammedia-Casablanca, Faculty of Science and Technology Mohammedia, BP 146 Mohammedia 20650, Morocco; Regional Laboratory of Analysis and Research of Oujda, National Office for the Safety of Food Products, 60 000 Oujda, Morocco.

Military Hospital of Instruction Mohammed V, Rabat 14 000, Morocco.

出版信息

J Virol Methods. 2014 Aug;204:38-43. doi: 10.1016/j.jviromet.2014.03.019. Epub 2014 Mar 31.

DOI:10.1016/j.jviromet.2014.03.019

PMID:24698762

Abstract

Sheeppox is now enzootic in Morocco. The development of a reliable method for rapid diagnosis of the disease is a central part of any control strategy. The aim of this study is to determine the diagnostic value of a variety of clinical samples such as ovine nasal, ocular or rectal swabs for the detection of sheeppox virus (SPPV) by qualitative conventional polymerase chain reaction (PCR), using a single pair of primers targeting the inverted terminal repeats of the SPPV InS-1 strain, a virulent field isolate. Swab and blood samples were collected from forty animals naturally infected with SPPV who had clinical signs of sheeppox. All animals tested PCR-positive for SPPV. Positive results were obtained infrequently with blood samples, whereas swab samples from at least two sites (nasal, ocular, rectal) were positive per evaluated animal. These results indicate that swab samples are suitable for quantitative molecular SPPV diagnosis. PCR product sequences obtained from all types of sheep samples proved to be identical to the corresponding regions of sheeppox virus strain Romania 65.

摘要

绵羊痘目前在摩洛哥呈地方流行性。开发一种可靠的疾病快速诊断方法是任何控制策略的核心部分。本研究的目的是通过定性常规聚合酶链反应（PCR），使用一对靶向绵羊痘病毒（SPPV）InS-1毒株（一种强毒力的野外分离株）反向末端重复序列的引物，确定各种临床样本（如绵羊鼻拭子、眼拭子或直肠拭子）对检测绵羊痘病毒（SPPV）的诊断价值。从40只自然感染SPPV且有绵羊痘临床症状的动物身上采集拭子和血液样本。所有动物的SPPV PCR检测均为阳性。血液样本很少获得阳性结果，而每只评估动物至少两个部位（鼻、眼、直肠）的拭子样本呈阳性。这些结果表明拭子样本适用于SPPV的定量分子诊断。从所有类型绵羊样本中获得的PCR产物序列被证明与绵羊痘病毒罗马尼亚65株的相应区域相同。