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瑞士流产绵羊临床条件下绵羊流产沙门氏菌感染的培养和PCR诊断

Diagnosis by culture and PCR of Salmonella abortusovis infection under clinical conditions in aborting sheep in Switzerland.

作者信息

Belloy Luc, Decrausaz Loane, Boujon Patrick, Hächler Herbert, Waldvogel Andreas S

机构信息

Institut Galli-Valerio, Lausanne, Switzerland.

出版信息

Vet Microbiol. 2009 Sep 18;138(3-4):373-7. doi: 10.1016/j.vetmic.2009.03.026. Epub 2009 Mar 25.

Abstract

Since 2003 eleven Swiss sheep flocks were affected by abortion storms due to Salmonella abortusovis, an infection which had not been reported in this country for decades although cases of salmonellosis are notifiable in Switzerland. This raised doubts about the adequacy of the currently used diagnostic tools and the origin of this infection. Therefore, PCR was tested for its potential as a more rapid and more reliable method for diagnosing S. abortusovis infections under field conditions. Fecal and vaginal samples were collected at different times after abortion and PCR was used to detect bacterial DNA. Bacteria were isolated by conventional culture techniques. For determining their origin they were analyzed by pulsed field gel electrophoresis (PFGE) and compared to isolates from Germany and France. Sequencing of randomly selected amplicons allowed confirming the specificity of the result. PCR was more sensitive because it allowed detecting S. abortusovis DNA up to three months after infection even in samples that were negative by culture. Escherichia coli from the digestive tract of sheep could inhibit the growth of S. abortusovis in vitro suggesting that the lower sensitivity of diagnosis by bacterial culture may in part be due to growth inhibition of S. abortusovis by resident bacteria. Results of PFGE indicated that the Swiss strains were closely related among themselves but distinct from German and French strains suggesting the presence of an autochthonous infection.

摘要

自2003年以来,瑞士有11个绵羊群因绵羊流产沙门氏菌而受到流产风暴的影响,尽管瑞士法定报告沙门氏菌病病例,但这种感染在该国已有数十年未被报告。这引发了人们对当前使用的诊断工具的充分性以及这种感染来源的质疑。因此,对聚合酶链反应(PCR)作为一种在野外条件下诊断绵羊流产沙门氏菌感染的更快速、更可靠方法的潜力进行了测试。在流产后的不同时间采集粪便和阴道样本,并使用PCR检测细菌DNA。通过传统培养技术分离细菌。为了确定它们的来源,通过脉冲场凝胶电泳(PFGE)对其进行分析,并与来自德国和法国的分离株进行比较。对随机选择的扩增子进行测序可确认结果的特异性。PCR更敏感,因为即使在培养呈阴性的样本中它也能在感染后长达三个月检测到绵羊流产沙门氏菌DNA。绵羊消化道中的大肠杆菌在体外可抑制绵羊流产沙门氏菌的生长,这表明细菌培养诊断敏感性较低可能部分归因于常驻细菌对绵羊流产沙门氏菌的生长抑制。PFGE结果表明,瑞士菌株彼此密切相关,但与德国和法国菌株不同,这表明存在本地感染。

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