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

F. Brunello, M. Ligozzi, E. Cristelli, S. Bonora, E. Tortoli, R. Fontana

Research output: Contribution to journalArticlepeer-review

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.

Original languageEnglish
Pages (from-to)2799-2806
Number of pages8
JournalJournal of Clinical Microbiology
Volume39
Issue number8
DOIs
Publication statusPublished - 2001

ASJC Scopus subject areas

  • Microbiology (medical)
  • Microbiology

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