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Abbreviations: aerobactin (iucD), afimbrial adhesin (afaB/C), cytolethal distending toxin (cdtB), cytotoxic necrotizing factor type 1 (cnf-1), G adhesin classes of P fimbriae (papG alleles), group II capsule (kpsMTII), α-hemolysin (hly), minor structural subunits of P fimbriae (papE/F), outer membrane protein of P fimbrae (papC), polymerase chain reaction (PCR), S fimbriae (sfaC/D), type 1 fimbriae (fimH), urinary tract infection (UTI), uropathogenic specific protein (usp), uropathogenic Escherichia coli (UPEC), virulence factors (VFs SUMMARYAdhesins (P-fimbriae, S-fimbriae, type 1 fimbriae and afimbrial adhesin), toxins (α-hemolysin and cytotoxic necrotizing factor type 1), iron acquisition systems (aerobactin) and host defense avoidance mechanisms (capsule or lipopolysaccharide) have been shown to be prevalent in Escherichia coli strains associated with urinary tract infections. In this work, 162 Uropathogenic Escherichia coli (UPEC) strains from patients with cystitis were genotypically characterized by polymerase chain reaction (PCR) assay. We developed three multiplex PCR assays for virulence-related genes papC, papE /F, papG alleles, fimH, sfa/foc, afaE, hly, usp, cdtB, iucD, and kpsMTII, all

The population structure of 71 carbapenem-resistant Acinetobacter baumannii clinical isolates from several hospitals in Brazil was investigated by ApaI pulsed-field gel electrophoresis, bla OXA-51 -like subtyping, and multilocus sequence typing (Institute Pasteur scheme). In addition to the predominance of strains carrying bla OXA-23 , we detected the presence of bla OXA-72 and bla OXA-231 . We observed a predominance of clonal complex 1 (CC1), CC15, and CC79 and representative strains of the worldwide-disseminated international clone I. The epidemiology of carbapenem-resistant Acinetobacter baumannii (CRAB) strains indicates the predominance of major lineages around the world (1), reinforcing the need to monitor their spread. We evaluated the population structure of CRAB strains circulating in different hospitals in Brazil.Seventy-one CRAB strains, recovered from clinical specimens of nonrepetitive patients who attended 1 of 64 different health institutions located in 23 different cities across the state of São Paulo, Brazil, and were referred to Instituto Adolfo Lutz (a public health laboratory), were studied. Each strain was definitively identified as A. baumannii species (2), and resistances to imipenem and meropenem (Sigma-Aldrich, St. Louis, MO) were confirmed by the broth microdilution technique (3); the MIC 50 and MIC 90 were 16 and 32 g/ml, respectively, for both drugs. bla OXA group genes were detected by multiplex PCR (4, 5) and fully sequenced (6-8). The bla OXA-23 gene was detected in 68 strains (95.8%), and all of them were associated with ISAba1 upstream (9). The frequencies of bla and bla OXA-231 were 2.8% (two strains) and 1.4% (one strain), respectively; none of them were associated with upstream ISAba1. The bla OXA-58 -like gene was not detected. The bla KPC , bla NDM , bla SPM , bla VIM , and bla OXA-48 genes (10) were also not detected. Sequencing of the bla OXA-51 -like gene (11) revealed the predominance of the bla OXA-51 allele in 23 strains (32.4%), of bla OXA-69 in 21 strains (29.6%), and of bla OXA-65 in 21 strains (29.6%); three strains (4.2%) presented the bla OXA-64 allele, two (4.2%) presented the bla OXA-70 allele, and only one (1.4%) presented the bla OXA-88 variant. Pulsed-field gel electrophoresis (PFGE) (12) revealed the occurrence of 64 restriction profiles that were grouped into seven clusters (A to G) on the basis of a 70% cutoff. Multilocus sequence typing (MLST; Institute Pasteur scheme [see http://pubmlst.org/abaumannii/]) and the eBURST algorithm (http://eburst.mlst.net/) identified the predominance of three major clonal complexes (CC1, CC15, and CC79) comprising more than 90% (Table 1) of the CRAB strains circulating in different hospitals across the state of São Paulo in recent years. Considering that CC1, CC15, and CC79 in isolates of OXA-23-producing A. baumannii strains circulating in other regions of Brazil have already been described (14-18), we found evidence for the role of a limited number of clonal complexes of CRAB circulating in this subcontinental country...

Identification of Lactobacillus sp. strains by phenotypic methods may lead to doubtful results possibly interfering in the reliability of the epidemiological and probiotics studies. Therefore this study aimed to determine the best methodology for the identification of the large diversity of lactobacilli species found in the vagina by comparing two techniques, one based on their biochemical profile and other employing molecular biology. A carbohydrate fermentation test (API 50 CH) was compared with multiplex polymerase chain reaction (PCR) for the identification of species of vaginal lactobacilli from 135 healthy women. The kappa index was used to evaluate agreement between the methods. Using the molecular technique, L. crispatus (32.6%), L. jensenii (25%) and L. gasseri (20.6%) were the most frequent species. However, using the biochemical technique, the most frequent species were: L. acidophilus (34.8%), L. crispatus (27.2%) and L. fermentum (13%). Although L. acidophilus was the most frequent specie found by biochemical tests, no strain of this microorganism was detected by PCR. Agreement between the methods was low for identification of all the most common species. Although rates of L. crispatus detected were similar using both methods (32.6% and 27.2%), agreement between them was relatively low (kappa = 0.52). Conclusions: Our results confirmed the limitation of the biochemical method and the applicability of a previously published molecular method (Multiplex PCR) for the identification of lactobacilli in the vaginal tract, focusing on further necessity of its improvement for also targeting L. vaginalis and L. iners.

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