A multiplex-PCR for the differentiation of Mycobacterium bovis and Mycobacterium tuberculosis.

Abstract:

:A multiplex-polymerase chain reaction (PCR) assay based on one-step amplification and detection of two different mycobacterial genomic fragments was designed for differentiation of Mycobacterium bovis and Mycobacterium tuberculosis. The oligonucleotide primers were chosen from a 500-bp genomic fragment which is well conserved in M. bovis and the pncA gene (based on M. tuberculosis-specific nucleotide polymorphism, a cytosine residue at position 169), specific for M. tuberculosis. The multiplex-PCR allowed detection of a single product of 500 bp in M. bovis isolates while M. tuberculosis isolates generated a single product of 185 bp, with or without an additional product of 500 bp. None of the atypical mycobacterial isolates revealed any amplification products. The method was found to be highly specific and could detect as little as 20 pg of pure DNA. This multiplex-PCR assay, based on the 500-bp fragment and the pncA gene, may be very useful for the rapid and specific differentiation of these two closely related mycobacteria and easy to use in medical and veterinary microbiological laboratories.

journal_name

FEMS Microbiol Lett

authors

Shah DH,Verma R,Bakshi CS,Singh RK

doi

10.1111/j.1574-6968.2002.tb11322.x

keywords:

subject

Has Abstract

pub_date

2002-08-27 00:00:00

pages

39-43

issue

1

eissn

0378-1097

issn

1574-6968

pii

S0378109702008443

journal_volume

214

pub_type

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