Detection and identification of Mycobacterium tuberculosis, M. bovis/BCG, and M. avium by two-step polymerase chain reaction. Comparison with ELISA using A60 antigen

C Bollet; X De Lamballerie; C Zandotti; C Vignoli; M J Gevaudan; P De Micco

Detection and identification of Mycobacterium tuberculosis, M. bovis/BCG, and M. avium by two-step polymerase chain reaction. Comparison with ELISA using A60 antigen

Microbiologica. 1992 Oct;15(4):345-9.

Authors

C Bollet¹, X De Lamballerie, C Zandotti, C Vignoli, M J Gevaudan, P De Micco

Affiliation

¹ Laboratoire de Microbiologie et d'Hygiène Hospitalière, Hôpital Salvator, Marseille, France.

PMID: 1435347

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.

Publication types

Comparative Study

MeSH terms

Antigens, Bacterial / genetics*
Base Sequence
DNA, Bacterial / genetics*
DNA, Single-Stranded
Enzyme-Linked Immunosorbent Assay
Molecular Sequence Data
Mycobacterium / genetics
Mycobacterium / immunology
Mycobacterium / isolation & purification*
Mycobacterium avium / isolation & purification
Mycobacterium bovis / isolation & purification
Mycobacterium tuberculosis / isolation & purification
Polymerase Chain Reaction / methods*
Sensitivity and Specificity

Substances

Antigens, Bacterial
DNA, Bacterial
DNA, Single-Stranded