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粪便定量实时聚合酶链反应检测保育猪胞内劳森菌相关性增生性肠炎的诊断性能

Diagnostic performance of fecal quantitative real-time polymerase chain reaction for detection of Lawsonia intracellularis-associated proliferative enteropathy in nursery pigs.

作者信息

Pedersen Ken Steen, Stege Helle, Jensen Tim K, Guedes Roberto, Ståhl Marie, Nielsen Jens Peter, Hjulsager Charlotte, Larsen Lars E, Angen Øystein

机构信息

HERD-Centre for Herd Oriented Education, Research and Development, Department of Large Animal Sciences, University of Copenhagen, Groennegaardsvej 2, DK-1870 Frederiksberg C, Denmark.

出版信息

J Vet Diagn Invest. 2013 May;25(3):336-40. doi: 10.1177/1040638713480499. Epub 2013 Mar 27.

Abstract

Quantitative polymerase chain reaction (qPCR) tests for detection and quantification of Lawsonia intracellularis in feces from pigs have been developed. The objective of the current study was to evaluate the diagnostic performance of a fecal qPCR test for detection of nursery pigs with L. intracellularis-associated proliferative enteropathy (PE) under field conditions. Furthermore, the diagnostic performance for different subpopulations of pigs was investigated, including pigs infected or noninfected with Porcine circovirus-2, Brachyspira pilosicoli, and Escherichia coli. The diagnostic performance was evaluated in terms of diagnostic sensitivity and specificity. Data from pigs originating from 20 herds with antibiotic treatment requiring diarrhea outbreaks from a prior study were reused. Before treatment, pigs were randomly selected for histological and immunohistochemical examination of intestinal segments and fecal quantification of L. intracellularis by qPCR. A total of 313 pigs (157 without diarrhea, 156 with diarrhea) were included in the statistical analysis, and 37 pigs (11.8%) were classified as PE positives (defined as proliferative histological lesions in combination with L. intracellularis demonstration by immunohistochemistry). Lawsonia intracellularis was detected by qPCR in feces from 91 pigs (29.1%). A nonparametric receiver operating characteristic (ROC) analysis provided an area under the ROC curve (0.93) and an optimal cutoff value of 4.8 log10 L. intracellularis bacteria/g feces. This cutoff provided a diagnostic sensitivity of 0.84 and diagnostic specificity of 0.93. The diagnostic sensitivity and specificity were significantly different between herds (P < 0.0001). Numerically, diagnostic sensitivity and specificity were different between subpopulations of pigs, but no significant differences were demonstrated.

摘要

已经开发出用于检测和定量猪粪便中胞内劳森菌的定量聚合酶链反应(qPCR)检测方法。本研究的目的是评估粪便qPCR检测在现场条件下对检测患有胞内劳森菌相关增生性肠炎(PE)的保育猪的诊断性能。此外,还研究了不同猪亚群的诊断性能,包括感染或未感染猪圆环病毒2型、毛样短螺旋体和大肠杆菌的猪。从诊断敏感性和特异性方面评估了诊断性能。重复使用了来自先前一项研究中20个有抗生素治疗需求的腹泻暴发猪群的猪的数据。在治疗前,随机选择猪进行肠段的组织学和免疫组织化学检查以及通过qPCR对粪便中的胞内劳森菌进行定量。共有313头猪(157头无腹泻,156头有腹泻)纳入统计分析,37头猪(11.8%)被分类为PE阳性(定义为增生性组织学病变并通过免疫组织化学证明有胞内劳森菌)。通过qPCR在91头猪(29.1%)的粪便中检测到了胞内劳森菌。非参数受试者工作特征(ROC)分析得出ROC曲线下面积为0.93,最佳临界值为每克粪便4.8 log10胞内劳森菌。该临界值的诊断敏感性为0.84,诊断特异性为0.93。各猪群之间的诊断敏感性和特异性有显著差异(P < 0.0001)。从数值上看,不同猪亚群之间的诊断敏感性和特异性有所不同,但未显示出显著差异。

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