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口腔液样本实时 PCR 检测在筛选猪群中流行的胸膜肺炎放线杆菌血清型中的应用评估。

Assessment of real-time PCR test in oral fluid samples for screening the serotypes of Actinobacillus pleuropneumoniae circulating in swine herds.

机构信息

Exopol, Pol. Río Gállego D14. San Mateo de Gállego, Zaragoza, Spain.

Research Group on Infectious Diseases in Production Animals and Swine and Poultry Infectious Diseases Research Centre, Faculty of Veterinary Medicine, University of Montreal, 3200 Sicotte, Saint-Hyacinthe, Quebec J2S 2M2, Canada.

出版信息

Vet Microbiol. 2024 Nov;298:110268. doi: 10.1016/j.vetmic.2024.110268. Epub 2024 Sep 26.

Abstract

Actinobacillus pleuropneumoniae is the etiological agent of porcine pleuropneumonia, causing remarkable economic losses in the global swine industry. The diversity of A. pleuropneumoniae is generally determined through serotype identification, which is commonly employed for control strategies and surveillance. However, serological methods currently in use still have significant limitations. This study explores the use of real-time polymerase chain reaction (qPCR) to detect circulating serotypes of A. pleuropneumoniae in non-diseased swine herds through testing of oral fluids. The study included three A. pleuropneumoniae-positive and three A. pleuropneumoniae-negative farms located in Quebec, Canada. Tonsil brushings, microbiological growths, and oral fluids were analyzed using qPCR to detect A. pleuropneumoniae and its distinct serotypes. Serological tests were performed using the LPS ELISA available at that time. In negative farms the absence of A. pleuropneumoniae and any serotype confirmed the specificity of the method. Positive farms, on the other hand, confirmed also the sensitivity of the analysis, with oral fluid samples consistently yielding positive results for the serotypes identified by ELISA. The qPCR test conducted on oral fluids offers a noninvasive and cost-effective method for monitoring, complementing traditional serological techniques. It provides qualitative information about serotype distribution, facilitating proactive surveillance and control strategies.

摘要

胸膜肺炎放线杆菌是猪传染性胸膜肺炎的病原体,给全球养猪业造成了巨大的经济损失。胸膜肺炎放线杆菌的多样性通常通过血清型鉴定来确定,这在控制策略和监测中被广泛应用。然而,目前使用的血清学方法仍然存在很大的局限性。本研究探讨了通过检测口腔液,使用实时聚合酶链反应(qPCR)检测非发病猪群中循环的胸膜肺炎放线杆菌血清型的方法。该研究包括位于加拿大魁北克的三个胸膜肺炎放线杆菌阳性农场和三个胸膜肺炎放线杆菌阴性农场。使用 qPCR 对扁桃体刷检物、微生物生长物和口腔液进行分析,以检测胸膜肺炎放线杆菌及其不同的血清型。使用当时可用的 LPS ELISA 进行血清学检测。在阴性农场,没有检测到胸膜肺炎放线杆菌和任何血清型,这证实了该方法的特异性。阳性农场则证实了分析的敏感性,ELISA 鉴定的血清型的口腔液样本始终产生阳性结果。口腔液 qPCR 检测提供了一种非侵入性、具有成本效益的监测方法,补充了传统的血清学技术。它提供了关于血清型分布的定性信息,有助于主动监测和控制策略的制定。

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