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

Shah, D. H.; Verma, Rishendra; Bakshi, C.S. and Singh, R.K. (2002) A multiplex-PCR for the differentiation of Mycobacterium bovis and Mycobacterium tuberculosis. FEMS Microbiology Letters, 214 (1). pp. 39-43.

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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.

EPrint Type:Article
Uncontrolled Keywords:Mycobacterium bovis; Mycobacterium tuberculosis ; Multiplex-polymerase chain reaction
Subjects:Diagnosis > Diagnostic Techniques and Procedures
Bacterial Infections and Mycoses > Bacterial Infections > Gram-Positive Bacterial Infections > Actinomycetales Infections > Mycobacterium Infections > Tuberculosis
Bacterial Infections and Mycoses > Bacterial Infections > Gram-Positive Bacterial Infections > Actinomycetales Infections > Mycobacterium Infections
ID Code:81
Deposited By:Dr. Devendra Shah
Deposited On:26 July 2005

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