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Comparison of a semi-automated rep-PCR system and multilocus sequence typing for differentiation of Salmonella enterica isolates
Authors:Elloulu Ben-Darif  E John Threlfall  Mathew Upton
Institution:a Department of Medical Microbiology, School of Medicine, University of Manchester, Clinical Sciences Building, Manchester Royal Infirmary, Oxford Road, Manchester, M13 9WL, United Kingdom
b Health Protection Agency, Gastrointestinal, Emerging and Zoonotic Infections, Centre for Infections, 61 Colindale Avenue, London, NW9 5EQ, United Kingdom
c Health Protection Agency North West Laboratory, Clinical Science Building, Manchester Royal Infirmary, Manchester, M13 9WZ, United Kingdom
d Health Protection Agency, Food, Water and Environmental Microbiology Network, Preston Laboratory, Royal Preston Hospital, Sharoe Green Lane, Preston, PR2 9HT, United Kingdom
Abstract:The accurate sub-typing of Salmonella enterica isolates is essential for epidemiological investigations and surveillance of Salmonella infections. Salmonella isolates are currently identified using the Kauffman-White serotyping scheme. Multilocus sequence typing (MLST) schemes have been developed for the major bacterial pathogens including Salmonella and have assisted in understanding the molecular epidemiology and population biology of these organisms. Recently, the DiversiLab rep-PCR system has been developed using micro-fluidic chips to provide standardized, semi-automated fingerprinting for pathogens including S. enterica. In the current study, 71 isolates of S. enterica, representing 21 serovars, were analyzed using MLST and the DiversiLab rep-PCR system. MLST was able to identify 31 sequence types (STs), while the DiversiLab system revealed 38 DiversiLab types (DTs). The rep-PCR distinguished isolates of different serovars and showed greater discriminatory power (0.95) than MLST typing (0.89). Rep-PCR exhibited 92% concordance with MLST and 90% with serotyping, while the concordance level of MLST typing with serotyping was 96%, representing a strong association. Comparison of rep-PCR profiles with those held in an online library database led to the accurate prediction of serovar in 63% of cases and resulted in inaccurate predictions for 10% of profiles. MLST and the rep-PCR system may provide useful additional informative techniques for the molecular identification of S. enterica. We conclude that the DiversiLab rep-PCR system may provide a rapid (less than 4 h) and standardized method for sub-typing isolates of S. enterica.
Keywords:Diagnostics  Typing  MLST  Salmonella  DiversiLab
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