RESUMO
The new epidemic serovar O139 of Vibrio cholerae has emerged from the pandemic serovar O1 biotype El Tor through the replacement of a 22-kbp DNA region by a 40-kbp O139-specific DNA fragment. This O139-specific DNA fragment contains an insertion sequence that was described previously (U. H. Stroeher, K. E. Jedani, B. K. Dredge, R. Morona, M. H. Brown, L. E. Karageorgos, J. M. Albert, and P. A. Manning, Proc. Natl. Acad. Sci. USA 92:10374-10378, 1995) and designated IS1358O139. We studied the distribution of the IS1358 element in strains from various serovars by Southern analysis. Its presence was detected in strains from serovars O1, O2, O22, O139, and O155 but not in strains from serovars O15, O39, and O141. Furthermore, IS1358 was present in multiple copies in strains from serovars O2, O22, and O155. We cloned and sequenced four copies of IS1358 from V. cholerae O22 and one copy from V. cholerae O155. A comparison of their nucleotide sequences with those of O1 and O139 showed that they were almost identical. We constructed a transposon consisting of a kanamycin resistance gene flanked by two directly oriented copies of IS1358 to study the functionality of this element. Transposition of this element from a nonmobilizable plasmid onto the conjugative plasmid pOX38-Gen was detected in an Escherichia coli recA donor at a frequency of 1.2 x 10(-8). Sequence analysis revealed that IS1358 duplicates 10 bp at its insertion site.
Assuntos
Elementos de DNA Transponíveis/genética , DNA Bacteriano/genética , Vibrio cholerae/classificação , Vibrio cholerae/genética , Sequência de Bases , Cólera/epidemiologia , Cólera/microbiologia , Conjugação Genética , Primers do DNA/genética , Surtos de Doenças , Genes Bacterianos , Humanos , Resistência a Canamicina/genética , Dados de Sequência Molecular , Plasmídeos/genética , Sequências Repetitivas de Ácido Nucleico , Mapeamento por Restrição , Sorotipagem , Vibrio cholerae/patogenicidadeRESUMO
The new epidemic strain O139 of Vibrio cholerae, the etiologic agent of cholera, has probably emerged from the pandemic strain O1 E1 Tor through a genetic rearrangement involving the horizontal transfer of exogenous O-antigen- and capsule-encoding genes of unknown origin. In V. cholerae O139, these genes are associated with an insertion sequence designated IS1358O139. In this work, we studied the distribution of seven genes flanking the IS1358O139 element in 13 serovars of V. cholerae strains. All these O139 genes and an IS1358 element designated IS1358O22-1 were only found in V. cholerae O22 with a similar genetic organization. Sequence analysis of a 4.5-kb fragment containing IS1358O22-1 and the adjacent genes revealed that these genes are highly homologous to those of V. cholerae O139. These results suggest that strains of V. cholerae O22 from the environment might have been the source of the exogenous DNA resulting in the emergence of the new epidemic strain O139.
Assuntos
DNA Bacteriano/genética , Genes Bacterianos/genética , Vibrio cholerae/genética , Sequência de Bases , Southern Blotting , Humanos , Dados de Sequência Molecular , Especificidade da Espécie , Vibrio cholerae/classificaçãoRESUMO
We analyzed the rfaD locus of the novel epidemic Vibrio cholerae strain O139, a putative region of rearrangement. This region includes 4 orfs in the same orientation. Two orfs, rfaD(O139) and orf2(O139) were almost identical to those described in V. cholerae O1. In contrast, the two other orfs upstream from rfaD(O139), designated orfA(O139) and orfB(O139), were absent from V. cholerae O1, but present in environmental strains of V. cholerae O22, O141 and O155. These results suggest that a chromosomal rearrangement might have occurred in the vicinity of rfaD in V. cholerae O1, resulting in the emergence of V. cholerae O139. The putative source of exogenous DNA might have been V. cholerae O22, O141 and O155.
Assuntos
Antígenos de Bactérias/genética , Carboidratos Epimerases/genética , Rearranjo Gênico , Antígenos O/imunologia , Vibrio cholerae/genética , Vibrio cholerae/imunologia , Proteínas de Bactérias/genética , Southern Blotting , Mapeamento Cromossômico , Clonagem Molecular , Cosmídeos , Proteínas de Escherichia coli , Biblioteca Gênica , Genes Bacterianos , Dados de Sequência Molecular , Mapeamento por Restrição , Análise de Sequência de DNARESUMO
Sixty vancomycin-resistant vanA mutant Enterococcus faecium (VRE) isolates, collected during a 40-month period from 48 patients hospitalized in a French Cancer Referral Center, were typed by using random amplified polymorphic DNA (RAPD), and the results were compared with those previously obtained by typing with SmaI pulsed-field gel electrophoresis (PFGE), which is currently recognized as the "gold standard." The discriminating power of RAPD typing, with seven primers and 11 combinations of primers, was tested on 18 strains, and only the most discriminating combination was further tested on the whole collection. We compared the epidemiological usefulness of RAPD typing of 60 clinical VRE isolates with that of SmaI PFGE typing. With primers AP4 and ERIC1R, RAPD generated 30 patterns versus the 36 patterns generated by SmaI PFGE. However, this did not hamper the epidemiologically correct clustering of 15 related strains and the detection of multiple colonization in nine patients. We conclude that this simple RAPD technique is well suited to the epidemiological typing of VRE and the monitoring of its nosocomial spread.
