Genetic diversity of O157:H7 and non-O157 verocytotoxigenic Escherichia coli from Argentina inferred from multiple-locus variable-number tandem repeat analysis (MLVA)

Bustamante, Ana Victoria; Sanso, Andrea Mariel; Lucchesi, Paula María Alejandra; Parma, A

doi:10.1016/j.ijmm.2009.08.020

Cited by 19 publications

(24 citation statements)

References 34 publications

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“…These were identical for all isolates, consistent with what was found by Miko et al (33). Though variations in locus CNV003 have been reported (11,33), most isolates presumable give no amplification product for CNV003 (11,29,33). Ideally, MLVA should be more discriminatory in order to thoroughly differentiate between E. coli isolates of serogroup O26:H11.…”

Section: Discussionsupporting

confidence: 85%

Potentially Human-Pathogenic Escherichia coli O26 in Norwegian Sheep Flocks

Sekse

Sunde

Lindstedt

et al. 2011

Appl Environ Microbiol

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A national survey of Escherichia coli O26 in Norwegian sheep flocks was conducted, using fecal samples to determine the prevalence. In total, 491 flocks were tested, and E. coli O26 was detected in 17.9% of the flocks. One hundred forty-two E. coli O26 isolates were examined for flagellar antigens (H typing) and four virulence genes, including stx and eae, to identify possible Shiga toxin-producing E. coli (STEC) and enteropathogenic E. coli (EPEC). Most isolates (129 out of 142) were identified as E. coli O26:H11. They possessed eae and may have potential as human pathogens, although only a small fraction were identified as STEC O26:H11, giving a prevalence in sheep flocks of only 0.8%. Correspondingly, the sheep flock prevalence of atypical EPEC (aEPEC) O26:H11 was surprisingly high (15.9%). The genetic relationship between the E. coli O26:H11 isolates was investigated by pulsed-field gel electrophoresis (PFGE) and multilocus variable number tandem repeat analysis (MLVA), identifying 63 distinct PFGE profiles and 22 MLVA profiles. Although the MLVA protocol was less discriminatory than PFGE and a few cases of disagreement were observed, comparison by partition mapping showed an overall good accordance between the two methods. A close relationship between a few isolates of aEPEC O26:H11 and STEC O26:H11 was identified, but all the E. coli O26:H11 isolates should be considered potentially pathogenic to humans. The present study consisted of a representative sampling of sheep flocks from all parts of Norway. This is the first large survey of sheep flocks focusing on E. coli O26 in general, including results of STEC, aEPEC, and nonpathogenic isolates.

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Section: Discussionsupporting

confidence: 85%

Potentially Human-Pathogenic Escherichia coli O26 in Norwegian Sheep Flocks

Sekse

Sunde

Lindstedt

et al. 2011

Appl Environ Microbiol

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“…Interestingly, this resistance type has been reported previously to exhibit distinct PFGE-profiles within the 131 sequence type [26]. Notably, locus CVN014 is known to be the most variable locus in the GECM-10 assay [27], [28]. CVN016 also has been reported to exhibit variation among ESBL- E. coli isolates [10].…”

Section: Resultsmentioning

confidence: 81%

Sub-Typing of Extended-Spectrum-β-Lactamase-Producing Isolates from a Nosocomial Outbreak: Application of a 10-Loci Generic Escherichia coli Multi-Locus Variable Number Tandem Repeat Analysis

et al. 2013

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Extended-spectrum β-lactamase producing Escherichia coli (ESBL-E. coli) were isolated from infants hospitalized in a neonatal, post-surgery ward during a four-month-long nosocomial outbreak and six-month follow-up period. A multi-locus variable number tandem repeat analysis (MLVA), using 10 loci (GECM-10), for ‘generic’ (i.e., non-STEC) E. coli was applied for sub-species-level (i.e., sub-typing) delineation and characterization of the bacterial isolates. Ten distinct GECM-10 types were detected among 50 isolates, correlating with the types defined by pulsed-field gel electrophoresis (PFGE), which is recognized to be the ‘gold-standard’ method for clinical epidemiological analyses. Multi-locus sequence typing (MLST), multiplex PCR genotyping of bla CTX-M, bla TEM, bla OXA and bla SHV genes and antibiotic resistance profiling, as well as a PCR assay specific for detecting isolates of the pandemic O25b-ST131 strain, further characterized the outbreak isolates. Two clusters of isolates with distinct GECM-10 types (G06-04 and G07-02), corresponding to two major PFGE types and the MLST-based sequence types (STs) 131 and 1444, respectively, were confirmed to be responsible for the outbreak. The application of GECM-10 sub-typing provided reliable, rapid and cost-effective epidemiological characterizations of the ESBL-producing isolates from a nosocomial outbreak that correlated with and may be used to replace the laborious PFGE protocol for analyzing generic E. coli.

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“…Among non-O157:H7 serotypes, the loci which presented the lowest variability were CVN002, CVN007, CVN015, and CVN003. Furthermore, this last locus presented null alleles (no PCR amplification) in all isolates except for those belonging to O157:H7 and O145:NM serotypes (Bustamante et al, 2010). Similarly, Løbersli et al (2012) found this locus was absent in several serotypes and they only confirmed the presence of this locus among E. coli O145, O157, and O55:H7 isolates.…”

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confidence: 98%

“…Lately, our laboratory is focused on implementing multiple-locus variable-number tandem repeat analysis (MLVA) as genotyping method, which will be reviewed here. With regard to this objective we modified and adapted two previously described MLVA protocols, one exclusive for O157:H7 (Lindstedt et al, 2003; Keys et al, 2005; Noller et al, 2006), based on polymorphism in nine variable number of tandem repeats (VNTR) loci – named MLVA O157 in the present manuscript (Bustamante et al, 2009a), and the other, a generic E. coli assay (Lindstedt et al, 2007) based on seven VNTR loci – named MLVA G (Bustamante et al, 2010). …”

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confidence: 99%

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Subtyping of STEC by MLVA in Argentina

Bustamante¹,

Sanso²,

Parma³

et al. 2012

Front. Cell. Inf. Microbio.

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Shiga toxin-producing Escherichia coli (STEC) causes serious human illness such as hemolytic uremic syndrome (HUS). Argentina has the world’s highest rate of this syndrome, which is the leading cause of acute renal failure among children. E. coli O157:H7 is the most common cause of HUS, but a substantial and growing proportion of this illness is caused by infection due to non-O157 strains. Multiple-locus variable-number tandem repeat analysis (MLVA) has become an established technique to subtype STEC. This review will address the use of routine STEC subtyping by MLVA in order to type this group of isolates and to get insight into the genetic diversity of native STEC. With regard to these objectives we modified and adapted two MLVA protocols, one exclusive for O157 and the other, a generic E. coli assay. A total of 202 STEC isolates, from different sources and corresponding to 20 serotypes, have been MLVA genotyped in our laboratory. In our experience, MLVA constitutes a very sensitive tool and enables us to perform an efficient STEC subtyping. The diversity found in many serotypes may be useful for future epidemiological studies of STEC clonality, applied to O157 as well as to non-O157 isolates.

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Genetic diversity of O157:H7 and non-O157 verocytotoxigenic Escherichia coli from Argentina inferred from multiple-locus variable-number tandem repeat analysis (MLVA)

Cited by 19 publications

References 34 publications

Potentially Human-Pathogenic Escherichia coli O26 in Norwegian Sheep Flocks

Potentially Human-Pathogenic Escherichia coli O26 in Norwegian Sheep Flocks

Sub-Typing of Extended-Spectrum-β-Lactamase-Producing Isolates from a Nosocomial Outbreak: Application of a 10-Loci Generic Escherichia coli Multi-Locus Variable Number Tandem Repeat Analysis

Subtyping of STEC by MLVA in Argentina

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