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  • Title: Identification of 54 mycobacterial species by PCR-restriction fragment length polymorphism analysis of the hsp65 gene.
    Author: Brunello F, Ligozzi M, Cristelli E, Bonora S, Tortoli E, Fontana R.
    Journal: J Clin Microbiol; 2001 Aug; 39(8):2799-806. PubMed ID: 11473995.
    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.
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