title: Detection and identification of Mycobacterium tuberculosis, M. bovis/BCG, and M. avium by two-step polymerase chain reaction. Comparison with ELISA using A60 antigen.
abstract: We propose a rapid two-step PCR to amplify a 767-bp sequence present in the gene coding for the 65-kD antigen of mycobacteria. The high G+C content (80%) permitted annealing to occur at 70 degrees C, enhancing the specificity. The amplified fragment contains a restriction site for differentiation between M. tuberculosis, M. bovis/BCG, and M. avium. Complete diagnosis can be achieved in less than four hours without labelled probe or nucleic acid transfer.
authors.affiliation: Laboratoire de Microbiologie et d'Hygiène Hospitalière, Hôpital Salvator, Marseille, France.
authors.forename: C
authors.initials: C
authors.lastname: Bollet
authors.forename: X
authors.initials: X
authors.lastname: De Lamballerie
authors.forename: C
authors.initials: C
authors.lastname: Zandotti
authors.forename: C
authors.initials: C
authors.lastname: Vignoli
authors.forename: M J
authors.initials: MJ
authors.lastname: Gevaudan
authors.forename: P
authors.initials: P
authors.lastname: De Micco
mesh_terms: D000942:Antigens, Bacterial / Q000235:genetics*; D001483:Base Sequence; D004269:DNA, Bacterial / Q000235:genetics*; D004277:DNA, Single-Stranded; D004797:Enzyme-Linked Immunosorbent Assay; D008969:Molecular Sequence Data; D009161:Mycobacterium / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D009162:Mycobacterium avium / Q000302:isolation & purification; D009163:Mycobacterium bovis / Q000302:isolation & purification; D009169:Mycobacterium tuberculosis / Q000302:isolation & purification; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
chemical_list: D000942:Antigens, Bacterial; D004269:DNA, Bacterial; D004277:DNA, Single-Stranded