Multicolor Melting Curve Analysis-Based Multilocus Melt Typing of Vibrio parahaemolyticus

摘要: Vibrio parahaemolyticus is the leading cause of seafood-borne gastroenteritis outbreaks. To track source these diseases in a timely manner, high throughput typing method critical. We hereby describe novel genotyping for V. parahaemolyticus, termed multilocus melt (MLMT), based on sequence (MLST). MLMT utilizes melting curve analysis to interrogate allelic types set informative single nucleotide polymorphisms (SNPs) derived from housekeeping genes used MLST. For each SNP, one type generates distinct Tm values, which are converted into binary code. Multiple SNPs thus generate series codes, forming (MT) corresponding with (ST) Using 12 SNPs, scheme could resolve 218 V.parahaemolyticus isolates 50 MTs 56 STs. The discriminatory power and MLST was similar Simpson’s index diversity 0.638 0.646, respectively. global (adjusted Rand = 0.982) directional congruence Wallace coefficient, MT→ST 0.965; ST→MT 1.000) between two approaches high. entire procedure be finished within 3 h negligible hands time real-time PCR machine. conclude that provides reliable efficient approach might also find use other pathogens.