Rapid identification of mycobacteria to species level by restriction fragment length polymorphism analysis of hsp65 gene.

Important note: Information in this article was accurate in 2000. The state of the art may have changed since the publication date.

Rapid identification of mycobacteria to species level by restriction fragment length polymorphism analysis of hsp65 gene.

Abstr Gen Meet Am Soc Microbiol. 1999 May 30-Jun 3;99:133 (abstract no. C-141). Unique Identifier : AIDSLINE AIDS/20712046
Koksalan K; Ergin A; Kocagoz T; Uzun M; Gurler B; Ang O; Gunalp A; Istanbul University Medical School, Department of Microbiology,; Ankara, Turkey.

Abstract: Classical identification tests for mycobacteria, based on culture and biochemical tests may take several weeks and frequently fail to produce a precise identification. PCR- restriction fragment length polymorphism analysis (PRA) offers an easy, rapid and inexpensive way of identification of mycobacteria to species level. We have obtained BstEII and HaeIII digestion patterns of the PCR amplified 65kD heat shock protein gene of 11 species of mycobacteria which did not take place, in earlier studies, in the algorithms that included 34 species of mycobacteria. It was possible to identify distinct PRA patterns for M. gadium, M. parafortuitum, M. duvalii, M. aichiense, M. rhodesiae, M. chitae, M. austroafricanum, M. chubuense, M. tokaiense which enabled unambiguous identification of these 9 species and differentiate from each other and the 34 species defined in previous studies. However M. gallinarum and M. diernhoferi produced PRA patterns similar to M. vaccae and M. terrae, respectively. The sizes of the restriction fragments defined, are approximations, since they are determined by electrophoresis. The lack of a suitable molecular weight marker and the sequence information for most of the species, makes sometimes difficult to differentiate species with similar PRA patterns. Sequencing the PCR product, used in identification, for all the known species and production of a molecular weight marker that contain fragments the same size as all restriction fragments, may simplify this very practical method and help it to be established in routine clinical microbiology laboratories.

Keywords: ABSTRACT Chaperonins/*GENETICS Polymerase Chain Reaction *Polymorphism, Restriction Fragment Length Species Specificity
000730
A0070888

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