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Background  Nontyphoidal  Salmonella  spp. constitute a major bacterial cause of food poisoning. Each  Salmonella  serotype causes distinct virulence to humans.    Method  A small cohort study was conducted to characterize several aspects of  Salmonella  isolates obtained from stool of diarrheal patients ( n  = 26) admitted to Phayao Ram Hospital, Phayao province, Thailand. A simple CRISPR 2 molecular analysis was developed to rapidly type  Salmonella  isolates employing both uniplex and high resolution melting (HRM) curve analysis.    Results  CRISPR 2 monoplex PCR generated a single  Salmonella  serotype-specific amplicon, showing  S.  4,[5],12:i:- with highest frequency (42%),  S.  Enteritidis (15%) and  S.  Stanley (11%);  S.  Typhimurium was not detected. CRISPR 2 HRM-PCR allowed further classification of  S.  4,[5],12:i:- isolates based on their specific CRISPR 2 signature sequences. The highest prevalence of  Salmonella  infection was during the summer season (April to August). Additional studies were conducted using standard multiplex HRM-PCR typing, which confirmed CRISPR 2 PCR results and, using a machine-learning algorithm, clustered the majority of  Salmonella  serotypes into six clades; repetitive element-based (ERIC) PCR, which clustered the serotypes into three clades only; antibiogram profiling, which revealed the majority resistant to ampicillin (69%); and test for extended spectrum  β -lactamase production (two isolates) and PCR-based detection of  bla  alleles.    Conclusion  CRISPR 2 PCR provided a simple assay for detection and identification of clinically-relevant  Salmonella  serotypes. In conjunction with antibiogram profiling and rapid assay for  β -lactamase producers, this approach should facilitate detection and appropriate treatment of Salmonellosis in a local hospital setting. In addition, CRISPR 2 HRM-PCR profiling enabled clustering of  S.  4,[5],12:i:-isolates according to CRISPR 2 locus signature sequences, indicative of their different evolutionary trajectories, thereby providing a powerful tool for future epidemiological studies of virulent  Salmonella  serotypes.

peerj

CRISPR 2 PCR and high resolution melting profiling for identification and characterization of clinically-relevant <i>Salmonella</i> enterica subsp. enterica

CRISPR 2 PCR and high resolution melting profiling for identification and characterization of clinically-relevant Salmonella enterica subsp. enterica