中国动物传染病学报2018，Vol.26Issue(1)：48-54,7.
检测创伤弧菌两种毒力基因的环介导等温扩增检测方法的建立
ESTABLISHMENT OF LOOP MEDIATED ISOTHERMAL AMPLIFICATION FOR DETECTION OF TWO VIRULENT GENES OF VIBRIO VULNIFICUS
摘要
Abstract
In the present study, we developed and validated a loop-mediated isothermal amplification (LAMP) assay for detection of two toxins genes of Vibrio vulnificus.The inner and outer primers of LAMP were designed based on the conserved sequences of V.vulnificus hemolysin gene A (HA) and repeats-in-toxin (RTX) virulent gene. The detection method was preliminary evaluated by formation of the white deposition and confirmed through agarose gel electrophoresis. The sensitivity of this detection method was 10 CFU/mL for two LAMP genes of V.vulnificus.This method showed good specificity to V.vulnificus as it did not react with any other bacterial species.The bacterial DNA was extracted by water-boiling method and it took 1.5 h from sample processing to obtaining results. The detection limits were 102CFU/g for RTX and 104CFU/g for HA. On the contrast, the detection limit of common PCR method was 103CFU/g for RTX and 104CFU/g for HA and it took 4 h to complete test.This LAMP method may be used for on-site diagnostics of V.vulnificus in the food and agricultural industries.关键词
环介导等温扩增/溶血素A基因/多重毒素/创伤弧菌Key words
Loop-mediated isothermal amplification/hemolysin gene A/repeats-in-toxin/Vibrio vulnificus分类
农业科技引用本文复制引用
别闯南,王权,白雪瑞,凌娇,李恭贺,蒋蔚..检测创伤弧菌两种毒力基因的环介导等温扩增检测方法的建立[J].中国动物传染病学报,2018,26(1):48-54,7.基金项目
上海市科学技术委员会科研计划项目(17140900400) （17140900400）
