[In silico analysis of the capability of two polimerase chain reaction techniques for stx gene detection].
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. Twenty-five 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.
Galli
L
L
Servicio Fisiopatogenia, Departamento Bacteriología, Instituto Nacional de Enfermedades Infecciosas, ANLIS Dr. Carlos G. Malbrán, Av. Vélez Sarsfield 563, 1281 Ciudad Autónoma de Buenos Aires, Argentina. lgalli@anlis.gov.ar
Leotta
G A
GA
Gugliada
M J
MJ
Rivas
M
M
D019295:Computational Biology; D016133:Polymerase Chain Reaction / Q000379:methods*; D022621:Shiga Toxin / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification*
D022621:Shiga Toxin