Revista argentina de microbiología
versión On-line ISSN 1851-7617
GALLI, L.; LEOTTA, G. A.; GUGLIADA, M. J. y RIVAS, M.. In silico analysis of the capability of two polimerase chain reaction techniques for stx gene detection. Rev. argent. microbiol. [online]. 2008, vol.40, n.1, pp. 9-12. ISSN 1851-7617.
Shiga toxin-producing Escherichia coli is an emergent pathogen, being the Shiga toxin (Stx) the main virulence factor. These toxins are classified into 6 types (1, 2, 2c, 2d, 2e and 2f) and 22 variants. In Argentina, two PCR for stx gene detection, PCR-MK and multiplex-PCR, were validated. The aim of this work was to analyze, by using bioinformatic tools, the stx variants that could be amplified by these PCRs, and to experimentally show the amplification of 8 stx variants. Twentyfive nucleotide sequences were collected from GenBank corresponding to 21 stx variants. The BLAST 2 sequences program was used to analyze the complementarities between the nucleotide sequence of the variants and the primers corresponding to the PCR studied. PCR-MK could detect types stx1, stx2, stx2c, stx2d and stx2f, but not type stx2e and three type stx2c variants. On the other hand, the multiplex-PCR could detect types stx1, stx2, stx2c, stx2d, but not stx2e and stx2f types. It was experimentally determined that both PCRs can detect those variants that cause severe disease in humans.
Palabras clave : Escherichia coli; Shiga toxin; Stx; PCR; Bioinformatics.