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

Citation
F. Brunello et al., Identification of 54 mycobacterial species by PCR-restriction fragment length polymorphism analysis of the hsp65 gene, J CLIN MICR, 39(8), 2001, pp. 2799-2806
Citations number
31
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Clinical Immunolgy & Infectious Disease",Microbiology
Journal title
JOURNAL OF CLINICAL MICROBIOLOGY
ISSN journal
0095-1137 → ACNP
Volume
39
Issue
8
Year of publication
2001
Pages
2799 - 2806
Database
ISI
SICI code
0095-1137(200108)39:8<2799:IO5MSB>2.0.ZU;2-X
Abstract
A total of 121 reference and clinical strains of both slowly and rapidly gr owing mycobacteria belonging to 54 species were studied for restriction fra gment length polymorphism of a PCR-amplified 439-bp segment of the gene enc oding 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 co mputer-aided comparison program. An algorithm describing these 54 species ( including 22 species not previously described) is proposed. We found that t his assay based on 10% PAGE provided a more precise estimate than that base d on agarose gel electrophoresis of the real size of restriction fragments as deduced from the sequence analysis and allowed identification of mycobac teria whose PCR-restriction fragment length polymorphism analysis patterns were unequivocally identified by fragments shorter than 60 bp.