Detection and identification of Mycobacterium tuberculosis, M. bovis/BCG, and M. avium by two-step polymerase chain reaction. Comparison with ELISA using A60 antigen.
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.
Bollet
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C
Laboratoire de Microbiologie et d'Hygiène Hospitalière, Hôpital Salvator, Marseille, France.
De Lamballerie
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X
Zandotti
C
C
Vignoli
C
C
Gevaudan
M J
MJ
De Micco
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P
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
D000942:Antigens, Bacterial; D004269:DNA, Bacterial; D004277:DNA, Single-Stranded