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利用口腔液进行细菌、核酸和抗体检测,提高红斑丹毒丝菌感染的生前诊断。

Improving ante mortem diagnosis of Erysipelothrix rhusiopathiae infection by use of oral fluids for bacterial, nucleic acid, and antibody detection.

机构信息

Department of Veterinary Diagnostic and Production Animal Medicine, College of Veterinary Medicine, Iowa State University, Ames, IA, USA.

出版信息

J Microbiol Methods. 2013 Feb 15;92(2):113-21. doi: 10.1016/j.mimet.2012.11.014. Epub 2012 Nov 29.

Abstract

Swine erysipelas is an economically important disease caused by Erysipelothrix rhusiopathiae. Pen-based collection of oral fluids has recently been utilized for monitoring infection dynamics in swine operations. The diagnostic performance of bacterial isolation, real-time PCR, and antibody detection by enzyme-linked immunosorbent assay (ELISA) and fluorescent microbead-based immunoassay (FMIA) methods were evaluated on pen-based oral fluid samples from pigs experimentally infected with E. rhusiopathiae (n=112) and from negative controls (n=32). While real-time PCR was a sensitive method with an overall detection rate of 100% (7/7 pens) one day post inoculation (dpi), E. rhusiopathiae was successfully isolated in only 28.6% (2/7 pens). Anti-Erysipelothrix IgM and IgG antibodies in pen-based oral fluids were detected at 4 to 5 dpi by FMIA and at 5 and 8 dpi by ELISA. The number of infected animals per pen, and in particular the timing of antimicrobial treatment administration impacted bacterial isolation and ELISA results. In oral fluid field samples, E. rhusiopathiae DNA was found in 23.3% of the samples while anti-E. rhusiopathiae IgG and IgM antibodies were found in 59.6% and 5.5% of the samples, respectively. The results suggest that an algorithm integrating oral fluids as specimen and real-time PCR and FMIA as detection methods is effective for earlier detection of an erysipelas outbreak thereby allowing for a more effective treatment outcome.

摘要

猪丹毒是由红斑丹毒丝菌引起的一种具有重要经济意义的疾病。最近,已经利用基于笔的口腔液收集来监测猪养殖场的感染动态。在实验感染红斑丹毒丝菌的猪(n=112)和阴性对照(n=32)的基于笔的口腔液样本上,评估了细菌分离、实时 PCR 以及酶联免疫吸附测定(ELISA)和荧光微球免疫分析(FMIA)抗体检测的诊断性能。虽然实时 PCR 是一种敏感方法,总体检测率为 100%(7/7 个笔)接种后一天(dpi),但红斑丹毒丝菌仅成功分离出 28.6%(2/7 个笔)。通过 FMIA 在 4 至 5 dpi 检测到基于笔的口腔液中的抗红斑丹毒丝菌 IgM 和 IgG 抗体,通过 ELISA 在 5 和 8 dpi 检测到。每支笔的感染动物数量,特别是抗菌治疗的时间,都会影响细菌分离和 ELISA 的结果。在口腔液现场样本中,发现 23.3%的样本中存在红斑丹毒丝菌 DNA,而 59.6%和 5.5%的样本中分别存在抗红斑丹毒丝菌 IgG 和 IgM 抗体。结果表明,整合口腔液作为标本,实时 PCR 和 FMIA 作为检测方法的算法,可有效早期检测丹毒爆发,从而获得更有效的治疗效果。

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