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J Clin Microbiol. 2001 Aug;39(8):2799-806.

Identification of 54 mycobacterial species by PCR-restriction fragment length polymorphism analysis of the hsp65 gene.

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1
Dipartimento di Patologia, Sezione di Microbiologia, Università di Verona, Italy.

Abstract

A total of 121 reference and clinical strains of both slowly and rapidly growing mycobacteria belonging to 54 species were studied for restriction fragment length polymorphism of a PCR-amplified 439-bp segment of the gene encoding the 65-kDa heat shock protein. Restriction digests were separated by 10% polyacrylamide gel electrophoresis (PAGE). By including a size standard in each sample, the restriction fragment profile was calculated using a computer-aided comparison program. An algorithm describing these 54 species (including 22 species not previously described) is proposed. We found that this assay based on 10% PAGE provided a more precise estimate than that based on agarose gel electrophoresis of the real size of restriction fragments as deduced from the sequence analysis and allowed identification of mycobacteria whose PCR-restriction fragment length polymorphism analysis patterns were unequivocally identified by fragments shorter than 60 bp.

PMID:
11473995
PMCID:
PMC88242
DOI:
10.1128/JCM.39.8.2799-2806.2001
[Indexed for MEDLINE]
Free PMC Article
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