caa a22 4500 605498520 CHVBK 20210128100546.0 cr unu---uuuuu 210128e20151001xx s 000 0 eng 10.1007/s00253-015-6831-7 doi (NATIONALLICENCE)springer-10.1007/s00253-015-6831-7 A multiplex oligonucleotide ligation-PCR as a complementary tool for subtyping of Salmonella Typhimurium [Elektronische Daten] [Véronique Wuyts, Wesley Mattheus, Nancy Roosens, Kathleen Marchal, Sophie Bertrand, Sigrid De Keersmaecker] Subtyping below the serovar level is essential for surveillance and outbreak detection and investigation of Salmonella enterica subsp. enterica serovar Typhimurium (S. Typhimurium) and its monophasic variant 1,4,[5],12:i:- (S. 1,4,[5],12:i:-), frequent causes of foodborne infections. In an attempt to overcome the intrinsic shortcomings of currently used subtyping techniques, a multiplex oligonucleotide ligation-PCR (MOL-PCR) assay was developed which combines different types of molecular markers in a high-throughput microsphere suspension array. The 52 molecular markers include prophage genes, amplified fragment length polymorphism (AFLP) elements, Salmonella genomic island 1 (SGI1), allantoinase gene allB, MLVA locus STTR10, antibiotic resistance genes, single nucleotide polymorphisms (SNPs) and phase 2 flagellar gene fljB. The in vitro stability of these markers was confirmed in a serial passage experiment. The validation of the MOL-PCR assay for subtyping of S. Typhimurium and S. 1,4,[5],12:i:- on 519 isolates shows that the method is rapid, reproducible, flexible, accessible, easy to use and relatively inexpensive. Additionally, a 100% typeability and a discriminatory power equivalent to that of phage typing were observed, and epidemiological concordance was assessed on isolates of 2 different outbreaks. Furthermore, a data analysis method is provided so that the MOL-PCR assay allows for objective, computerised data analysis and data interpretation of which the results can be easily exchanged between different laboratories in an international surveillance network. The Author(s), 2015 MOL-PCR nationallicence Salmonella Typhimurium nationallicence Subtyping nationallicence Luminex nationallicence Microsphere suspension array nationallicence Wuyts Véronique Department of Microbial and Molecular Systems, KU Leuven, Kasteelpark Arenberg 20 bus 2460, 3001, Leuven, Belgium aut Mattheus Wesley National Reference Centre for Salmonella and Shigella, Bacterial Diseases Division, Communicable and Infectious Diseases, Scientific Institute of Public Health (WIV-ISP), Juliette Wytmanstraat 14, 1050, Brussels, Belgium aut Roosens Nancy Platform Biotechnology and Molecular Biology, Scientific Institute of Public Health (WIV-ISP), Juliette Wytmanstraat 14, 1050, Brussels, Belgium aut Marchal Kathleen Department of Microbial and Molecular Systems, KU Leuven, Kasteelpark Arenberg 20 bus 2460, 3001, Leuven, Belgium aut Bertrand Sophie National Reference Centre for Salmonella and Shigella, Bacterial Diseases Division, Communicable and Infectious Diseases, Scientific Institute of Public Health (WIV-ISP), Juliette Wytmanstraat 14, 1050, Brussels, Belgium aut De Keersmaecker Sigrid Platform Biotechnology and Molecular Biology, Scientific Institute of Public Health (WIV-ISP), Juliette Wytmanstraat 14, 1050, Brussels, Belgium aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/19(2015-10-01), 8137-8149 0175-7598 99:19<8137 2015 99 253 https://doi.org/10.1007/s00253-015-6831-7 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-015-6831-7 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Wuyts Véronique Department of Microbial and Molecular Systems, KU Leuven, Kasteelpark Arenberg 20 bus 2460, 3001, Leuven, Belgium aut NATIONALLICENCE 700 1- Mattheus Wesley National Reference Centre for Salmonella and Shigella, Bacterial Diseases Division, Communicable and Infectious Diseases, Scientific Institute of Public Health (WIV-ISP), Juliette Wytmanstraat 14, 1050, Brussels, Belgium aut NATIONALLICENCE 700 1- Roosens Nancy Platform Biotechnology and Molecular Biology, Scientific Institute of Public Health (WIV-ISP), Juliette Wytmanstraat 14, 1050, Brussels, Belgium aut NATIONALLICENCE 700 1- Marchal Kathleen Department of Microbial and Molecular Systems, KU Leuven, Kasteelpark Arenberg 20 bus 2460, 3001, Leuven, Belgium aut NATIONALLICENCE 700 1- Bertrand Sophie National Reference Centre for Salmonella and Shigella, Bacterial Diseases Division, Communicable and Infectious Diseases, Scientific Institute of Public Health (WIV-ISP), Juliette Wytmanstraat 14, 1050, Brussels, Belgium aut NATIONALLICENCE 700 1- De Keersmaecker Sigrid Platform Biotechnology and Molecular Biology, Scientific Institute of Public Health (WIV-ISP), Juliette Wytmanstraat 14, 1050, Brussels, Belgium aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/19(2015-10-01), 8137-8149 0175-7598 99:19<8137 2015 99 253