A European interlaboratory trial to evaluate the performance of different PCR methods for Mycoplasma bovis diagnosis

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dc.contributor.author Wisselink, H.
dc.contributor.author Wisselink, H.
dc.contributor.author Smid, B.
dc.contributor.author Plater, J.
dc.contributor.author Ridley, A.
dc.contributor.author Andersson, A.-M.
dc.contributor.author Aspán, M.
dc.contributor.author Pohjanvirta, T.
dc.contributor.author Vähänikkilä, N.
dc.contributor.author Larsen, H.
dc.contributor.author Høgberg, J.
dc.contributor.author Colin, A.
dc.contributor.author Tardy, F.
dc.date.accessioned 2019-06-20T09:54:18Z
dc.date.available 2019-06-20T09:54:18Z
dc.date.issued 2019-03
dc.identifier.uri http://hdl.handle.net/10138/303381
dc.description.abstract Background Several species-specific PCR assays, based on a variety of target genes are currently used in the diagnosis of Mycoplasma bovis infections in cattle herds with respiratory diseases and/or mastitis. With this diversity of methods, and the development of new methods and formats, regular performance comparisons are required to ascertain diagnostic quality. The present study compares PCR methods that are currently used in six national veterinary institutes across Europe. Three different sample panels were compiled and analysed to assess the analytical specificity, analytical sensitivity and comparability of the different PCR methods. The results were also compared, when appropriate, to those obtained through isolation by culture. The sensitivity and comparability panels were composed of samples from bronchoalveolar fluids of veal calves, artificially contaminated or naturally infected, and hence the comparison of the different methods included the whole workflow from DNA extraction to PCR analysis. Results The participating laboratories used i) five different DNA extraction methods, ii) seven different real-time and/or end-point PCRs targeting four different genes and iii) six different real-time PCR platforms. Only one commercial kit was assessed; all other PCR assays were in-house tests adapted from published methods. The analytical specificity of the different PCR methods was comparable except for one laboratory where Mycoplasma agalactiae was tested positive. Frequently, weak-positive results with Ct values between 37 and 40 were obtained for non-target Mycoplasma strains. The limit of detection (LOD) varied from 10 to 103 CFU/ml to 103 and 106 CFU/ml for the real-time and end-point assays, respectively. Cultures were also shown to detect concentrations down to 102 CFU/ml. Although Ct values showed considerable variation with naturally infected samples, both between laboratories and tests, the final result interpretation of the samples (positive versus negative) was essentially the same between the different laboratories. Conclusion With a few exceptions, all methods used routinely in the participating laboratories showed comparable performance, which assures the quality of diagnosis, despite the multiplicity of the methods. fi
dc.language.iso en fi
dc.relation.ispartofseries BMC Veterinary Research 2019: Vol. 15, No. 86. fi
dc.subject Mycoplasma bovis fi
dc.subject Bovine respiratory disease fi
dc.subject Real-time PCR fi
dc.subject End-point PCR fi
dc.subject Culture fi
dc.subject Ring trial fi
dc.subject Bronchoalveolar lavage fluid fi
dc.title A European interlaboratory trial to evaluate the performance of different PCR methods for Mycoplasma bovis diagnosis fi
dc.type Artikkeli fi

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