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TITLE:  Comparison of multilocus sequence typing, pulsed-field gel electrophoresis, and antimicrobial susceptibility typing for characterization of Salmonella enterica serotype Newport isolates
AUTHORS:  Harbottle H;White DG;McDermott PF;Walker RD;Zhao S;
YEAR:  2006
JOURNAL ABBREV:  J Clin Microbiol
JOURNAL FULL:  Journal of clinical microbiology
END PAGE:  2457
KEYWORDS:  Ampicillin;Animals;Anti-Bacterial Agents;Bacterial Typing Techniques;Cattle;chemistry;Chickens;classification;Deoxyribonucleases,Type II Site-Specific;Dna;DNA Fingerprinting;DNA,Bacterial;drug effects;Drug Resistance,Multiple,Bacterial;Electrophoresis,Gel,Pulsed-Field;Food;Food Microbiology;Genes,Bacterial;genetics;Humans;isolation & purification;Kanamycin;Meat Products;Microbial Sensitivity Tests;microbiology;pharmacology;Phenotype;Polymorphism,Restriction Fragment Length;Public Health;Research;Salmonella;Salmonella enterica;Salmonella Infections;Salmonella Infections,Animal;Sequence Analysis,DNA;Serotyping;Streptomycin;Sulfamethoxazole;Swine;Tetracycline;Turkeys;United States;veterinary;Veterinary Medicine;
ABSTRACT:  In the United States, multidrug-resistant phenotypes of Salmonella enterica serotype Newport (commonly referred to as MDR-AmpC) have emerged in animals and humans and have become a major public health problem. Although pulsed-field gel electrophoresis (PFGE) is the current 'gold standard' typing method for Salmonella, multilocus sequence typing (MLST) may be more relevant to investigations exploring evolutionary and population biology relationships. In this study, 81 Salmonella enterica serotype Newport isolates from humans, food animals, and retail foods were examined for antimicrobial susceptibility and characterized using PFGE and MLST of seven genes, aroC, dnaN, hemD, hisD, purE, sucA, and thrA. Forty-nine percent of the isolates were resistant to nine or more of the tested antimicrobials. Salmonella isolates displayed resistance most often to sulfamethoxazole (57%), streptomycin (56%), tetracycline (56%), ampicillin (52%), and ceftiofur (49%) and, to a lesser extent, to kanamycin (19%), trimethoprim-sulfamethoxazole (17%), and gentamicin (11%). A total of 43 PFGE patterns were generated using XbaI, indicating a genetically diverse population. The largest PFGE cluster contained isolates from clinically ill swine, cattle, and humans. MLST resulted in 12 sequence types (STs), with one type encompassing 62% of the strains. Ten new sequence types and one novel allele type were identified. Furthermore, MLST typing showed that strains closely related by PFGE clustered in major STs, whereas more distantly related strains were separated into two clusters by PFGE. The results of this study demonstrated that the MLST scheme employed here clustered S. enterica serovar Newport isolates in distinct molecular populations, and strain discrimination was enhanced by combining PFGE, antimicrobial susceptibility, and MLST results
AFFILIATIONS:  Division of Animal and Food Microbiology, Office of Research, Center for Veterinary Medicine, U.S. Food and Drug Administration, 8401 Muirkirk Rd., Laurel, MD 20708, USA. heather.harbottle@fda.hhs.gov