In:
PeerJ, PeerJ, Vol. 8 ( 2020-06-16), p. e9113-
Abstract:
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.
Type of Medium:
Online Resource
ISSN:
2167-8359
DOI:
10.7717/peerj.9113/fig-1
DOI:
10.7717/peerj.9113/fig-2
DOI:
10.7717/peerj.9113/fig-3
DOI:
10.7717/peerj.9113/fig-4
DOI:
10.7717/peerj.9113/fig-5
DOI:
10.7717/peerj.9113/fig-6
DOI:
10.7717/peerj.9113/table-1
DOI:
10.7717/peerj.9113/table-2
DOI:
10.7717/peerj.9113/supp-1
DOI:
10.7717/peerj.9113/supp-2
Language:
English
Publisher:
PeerJ
Publication Date:
2020
detail.hit.zdb_id:
2703241-3
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