title: Development of one-tube multiplex polymerase chain reaction (PCR) for detecting Mycobacterium bovis.
abstract: A multiplex PCR (m-PCR) with primers targeting the 16S rRNA, Rv3873 and a 12.7-kb fragment in the genomes of a Mycobacterium tuberculosis complex was designed for the differential diagnosis of M. tuberculosis, M. bovis, M. bovis BCG and non-tuberculosis Mycobacterium (NTM). The specificity of this assay was 100%, and the detection limit was 15 pg of genomic DNA. Of the 206 blinded clinical samples, the detection rate of M. bovis infection by m-PCR was lower than that of the interferon gamma (IFN-γ) release assay; however, the false-positive rate by the tuberculin skin test and false-negative samples in the IFN-γ release assay were reduced. Our findings indicated that our m-PCR method is a useful tool for complementation to differentiate M. bovis from M. tuberculosis and NTM species.
authors.affiliation: Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, Jiangsu, China.
authors.forename: Zhang
authors.initials: Z
authors.lastname: Quan
authors.forename: Tan
authors.initials: T
authors.lastname: Haiming
authors.forename: Cai
authors.initials: C
authors.lastname: Xiaoyao
authors.forename: Yuan
authors.initials: Y
authors.lastname: Weifeng
authors.forename: Jia
authors.initials: J
authors.lastname: Hong
authors.forename: Zhu
authors.initials: Z
authors.lastname: Hongfei
mesh_terms: D000818:Animals; D002417:Cattle; D004269:DNA, Bacterial / Q000235:genetics; D060885:Multiplex Polymerase Chain Reaction / Q000662:veterinary*; D009161:Mycobacterium / Q000145:classification / Q000235:genetics / Q000302:isolation & purification; D009163:Mycobacterium bovis / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D012336:RNA, Ribosomal, 16S / Q000032:analysis; D012680:Sensitivity and Specificity
chemical_list: D004269:DNA, Bacterial; D012336:RNA, Ribosomal, 16S