Mycoplasma bovis real-time polymerase chain reaction assay validation and diagnostic performance

Mycoplasma bovis is an important bacterial pathogen in cattle, producing a variety of clinical diseases The organism, which requires specialized culture conditions and extended Incubation times to isolate and identify is frequently associated with concurrent infection with other pathogens which can potentially be more easily identified Real time polymerase chain reaction (real-time PCR) is a valuable diagnostic technique that can rapidly identify infectious agents in clinical specimens A real-time PCR assay was designed based on the uvrC gene to identify M bovis in diagnostic samples Using culture as the gold standard test, the assay performed well in a variety of diagnostic matrices Initial validation testing was conducted on 122 milk samples (sensitivity 88 9% [95% confidence interval (CI) 68 4-100%], specificity 100%), 154 lung tissues (sensitivity 89 0% [95% CI 83 1-94 9%], specificity 97 8% [95% CI 93 5-100%]), 70 joint tissue/fluid specimens (sensitivity 92 3% [95% CI 82 1-100%], specificity 95 5% [95% CI 89 3-100%]), and 26 nasal swabs (sensitivity 75 0% [95% CI 45 0-100%], specificity 83 3% [95% CI 66 1-100%]) Low numbers of other sample matrices showed good agreement between results of culture and PCR A review of clinical cases from 2009 revealed that, in general, PCR was used much more frequently than culture and provided useful diagnostic information in conjunction with clinical signs signalment and gross and histopathologic lesions Diagnostic performance of the real-time PCR assay developed as a testing method indicates that it is a rapid, accurate assay that is adaptable to a variety of PCR platforms and can provide reliable results on an array of clinical samples