high-resolution melting system to perform multilocus sequence typing of campylobacter jejuni茎高分辨率融化系统执行顺序输入的空肠弯曲杆菌.pdfVIP

High-Resolution Melting System to Perform Multilocus Sequence Typing of Campylobacter jejuni ´ 1 1 2 1 Simon Levesque , Sophie Michaud , Robert D. Arbeit , Eric H. Frost * ´ ´ ´ ´ ´ 1 Department of Microbiology and Infectious Diseases, Faculte de Medecine et des Sciences de la Sante de l’Universite de Sherbrooke, Sherbrooke, Quebec, Canada, 2 Infectious Diseases Section, Tufts University School of Medicine, Boston, Massachusetts, United States of America Abstract Multi-locus sequence typing (MLST) has emerged as the state-of-the-art method for resolving bacterial population genetics but it is expensive and time consuming. We evaluated the potential of high resolution melting (HRM) to identify known MLST alleles of Campylobacter jejuni at reduced cost and time. Each MLST locus was amplified in two or three sub fragments, which were analyzed by HRM. The approach was investigated using 47 C. jejuni isolates, previously characterized by classical MLST, representing isolates from diverse environmental, animal and clinical sources and including the six most prevalent sequence types (ST) and the most frequent alleles. HRM was then applied to a validation set of 84 additional C. jejuni isolates from chickens; 92% of the alleles were resolved in 35 hours of laboratory time and the cost of reagents per isolate was $20 compared with $100 for sequence-based typing. HRM has the potential to complement sequence-based methods for resolving SNPs and to facilitate a wide range of genotyping studies. ´ Citation: Levesque S, Michaud S, Arbeit RD, Frost EH (2011) High-Resolution Melting System to Perform Multilocus Sequence