Evaluation of different diagnostic methods for the detection of Mycobacterium avium subsp paratuberculosis in boot swabs and liquid manure samples

被引:20
作者
Hahn, Nathalie [1 ]
Failing, Klaus [2 ]
Eisenberg, Tobias [3 ]
Schlez, Karen [3 ]
Zschoeck, Peter-Michael [3 ]
Donat, Karsten [1 ]
Einax, Esra [1 ]
Koehler, Heike [4 ]
机构
[1] Thuringian Anim Dis Fund, Anim Hlth Serv, Jena, Thuringia, Germany
[2] Justus Liebig Univ, Unit Biomath & Data Proc, Giessen, Hesse, Germany
[3] Hessian State Lab, Dept Vet Med, Giessen, Hesse, Germany
[4] Friedrich Loeffler Inst, Fed Res Inst Anim Hlth, Inst Mol Pathogenesis, Jena, Thuringia, Germany
来源
BMC VETERINARY RESEARCH | 2017年 / 13卷
关键词
Cultural isolation; Diagnostic performance; DNA extraction; Environmental samples; qPCR; REAL-TIME PCR; POLYMERASE-CHAIN-REACTION; WITHIN-HERD PREVALENCE; BOVINE FECAL SAMPLES; ENVIRONMENTAL-SAMPLES; SSP PARATUBERCULOSIS; DAIRY HERDS; BACTERIAL CULTURE; MAP DETECTION; FECES;
D O I
10.1186/s12917-017-1173-6
中图分类号
S85 [动物医学(兽医学)];
学科分类号
0906 ;
摘要
Background: Environmental sampling based on boot swabs and/or liquid manure samples is an upcoming strategy for the identification of paratuberculosis (paraTB) positive herds, but only limited data are available regarding the diagnostic performance of molecular detection methods (qPCR) versus faecal culture (FC) for this purpose. In the present study, the test characteristics of two different qPCR protocols (A and B) and a standardized FC protocol, for the detection of Mycobacterium avium subsp. paratuberculosis in boot swabs and liquid manure samples were evaluated. Results: In 19 paraTB unsuspicious and 58 paraTB positive herds boot swabs and liquid manure were sampled simultaneously and analyzed in three different diagnostic laboratories. Using boot swabs and liquid manure, a substantial to excellent accordance was found between both qPCRs, for boot swabs also with culture, while for liquid manure the detection rate of culture was decreased after prolonged storage at -20 degrees C. The quantitative results of both qPCR methods correlated well for the same sample and also for boot swabs and liquid manure from the same herd. When cut-off threshold cycle (C-T-)-values were applied as recommended by the manufacturers, herd level specificity (Sp) of qPCR B was below 100% for boot swabs and for both qPCRs for liquid manure. A decreased herd level sensitivity was encountered after adjustment of Sp to 100% and re-calculation of the cut-off C-T-values. Conclusions: qPCR is equally suitable as bacterial culture for the detection of Mycobacterium avium subsp. paratuberculosis in boot swabs and liquid manure samples. Both matrices represent easily accessible composite environmental samples which can be tested with reliable results. The data encourage qPCR testing of composite environmental samples for paraTB herd diagnosis.
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页数:8
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