RAPD-PCR typing of Yersinia enterocolitica (Enterobacteriaceae) O:3 serotype strains isolated from pigs and humans

Abstract

Sixteen strains of Yersinia enterocolitica serotype O:3, isolated from apparently healthy pigs collected in Rio de Janeiro, and four human strains of serotypes O:4, O:5, O:6 and O:13 were analyzed by RAPD-PCR. The strains were grouped into five genotypic profiles according to the amplification patterns obtained with three random primers. Fifteen of the 16 pig strains had identical amplification patterns, which was named genotypic profile 1. The one different profile was named genotypic profile 2. Genotypic profile 1 was also exhibited by the O:6 human serotype strain. The O:4 and O:13 human serotype strains showed similar amplification profiles with two primers. However, the third primer induced a distinct profile in each strain. Therefore, these two strains were placed into genotypic profile 3 and 4, respectively. Each primer produced a completely different amplification profile in the O:5 human serotype strain; therefore, it was named genotypic profile 5. The presence or absence of plasmids in the strains studied did not affect the amplification results. These results show that genetic variations can exist within a serotype, and strains of different serotypes can exhibit the same amplification profile when compared using other primers.

Genetic Diversity Among Yersinia Enterocolitica Isolated From Sewage, Raw Milk and Packed Foods

“…Each 25 µl reaction mix contains 30 ng genomic DNA, 1 U Taq DNA polymerase, 1 X Taq DNA polymerase buffer (Chromous Biotech, Bangalore), 2.5 mM MgCl2, 400 µMdNTPs (Helini Biomolecules, India) and 20 pmol / µl primer. RAPD used for the study of diversity among Y. enterocolitica isolates was performed as described by Leal et al (1999). RAPD data were coded, analysed, all molecular markers were combined in a final analysis.…”

Molecular epidemiology ofYersinia enterocoliticainfections

“…However, a stable interlaboratory RAPD protocol may be obtained by careful optimization of technical procedures (Blixt et al, 2003). Some studies have been conducted to characterize Y. enterocolitica isolates with RAPD (Rasmussen et al, 1994;Odinot et al, 1995;Leal et al, 1999). This method allows discrimination between Y. enterocolitica isolates belonging to different bioserotypes and also, in some cases, between isolates belonging to the same bioserotype (Odinot et al, 1995;Leal et al, 1999).…”

Evaluation of a RAPD-based typing scheme in a molecular epidemiology study of Vibrio cholerae O1, Brazil

“…Under standardized reaction conditions, it gives a distinct pattern for each sample analysed, which can be compared with others (Makino et al. 1995; Leal et al. 1999; Radu et al.…”

“…It uses oligonucleotide primers with arbitrary sequences that can serve for the study of several bacterial species (Welsh and McClelland 1990;Williams et al 1990). Under standardized reaction conditions, it gives a distinct pattern for each sample analysed, which can be compared with others (Makino et al 1995;Leal et al 1999;Radu et al 1999;Pereira et al 2002). These characteristics could be useful in constructing a schematic pattern to be employed for the comparison and grouping of isolates of the same species.…”