Rapid and simple universal Escherichia coli genotyping method based on Multiple-Locus Variable-Number of Tandem Repeats Analysis using single-tube multiplex PCR and standard gel electrophoresis.

Rapid and simple universal Escherichia coli genotyping method based on Multiple-Locus Variable-Number of Tandem Repeats Analysis using single-tube multiplex PCR and standard gel electrophoresis. Appl Environ Microbiol. 2019 Jan 04;: Authors: Caméléna F, Birgy A, Smail Y, Courroux C, Mariani-Kurkdjian P, Le Hello S, Bonacorsi S, Bidet P Abstract We developed a multiplex PCR method based on Multiple-Locus Variable-number of tandem repeats (VNTR)-Analysis (MLVA) designed for rapid typing of E. coli and Shigella isolates The method amplifying seven VNTRs doesn't require sequencing capillary nor fluorescent dyes. Amplification products are simply loaded on a standard agarose gel for electrophoresis and banding patterns are analyzed visually. We evaluated the method on 220 strains belonging to different collections: ECOR collection (n=72), O1:K1 isolates causing neonatal meningitis (n=38), extended-spectrum beta-lactamase-producing fecal isolates belonging to the worldwide ST131 clone (n=38), Shiga-toxin producing E. coli (STEC) of serogroups O157:H7 (n=21) and O26 (n=16, 8 belonging to an outbreak), 27 Shigella isolates (22 S. sonnei including 5 epidemic strains) and 8 reference strains. The performances were compared to MLST, the DiversiLab automated REP-PCR, PFGE and whole genome sequencing. On the ECOR collection, we found 66 different profiles. Among the clonal group O1:K1, 14 different profiles were identified. For the 37 STECs, we...
Source: Applied and Environmental Microbiology - Category: Microbiology Authors: Tags: Appl Environ Microbiol Source Type: research