PMA⁃SRCA可视化检测海产品中的活副溶血性弧菌  

PMA⁃SRCA Visual Detection of Viable Vibrio parahaemolyticus in Seafood

作　　者：王宇航 张蕴哲[1] 杨倩 徐慧 马晓燕[1,4] 苑宁 张伟[1,5,6] WANG Yuhang;ZHANG Yunzhe;YANG Qian;XU Hui;MA Xiaoyan;YUAN Ning;ZHANG Wei(College of Food Science and Technology,Hebei Agricultural University,Baoding 071000,Hebei,China;School of Public Health,Hebei University,Baoding 071000,Hebei,China;College of Science and Technology,Hebei Agricultural University,Cangzhou 061100,Hebei,China;Laboratory of Food Biotechnology Innovation,Hebei Agricultural University,Baoding 071000,Hebei,China;College of Life Sciences,Hebei Agricultural University,Baoding 071000,Hebei,China;Hebei Provincial Key Laboratory of Analysis and Control for Zoonoses Microbial,Baoding 071000,Hebei,China)

机构地区：[1]河北农业大学食品科技学院,河北保定071000 [2]河北大学公共卫生学院,河北保定071000 [3]河北农业大学理工系,河北沧州061100 [4]河北农业大学食品生物技术创新工作室,河北保定071000 [5]河北农业大学生命科学学院,河北保定071000 [6]河北省人畜共患病原微生物分析与防控重点实验室,河北保定071000

出　　处：《食品研究与开发》2024年第18期197-203,共7页Food Research and Development

基　　金：国家自然科学基金项目(32172288、31371772);河北省自然科学基金重点项目(C2019204342);中央引导地方科技发展资金项目(216Z5501G、226Z5503G、236Z5502G);河北省外专百人计划(360⁃0803⁃JSN⁃3YGS);河北农业大学食品加工学科群经费资助(2021⁃06);河北省博士后科研项目(B2021005007);河北省重点研究开发计划项目(18275501D);河北省教育厅科研项目(QN2022073);河北省自然科学基金项目(C2019204284);河北省自然科学基金生物农业联合基金项目(C2023204200、C2023204241)。

摘　　要：为可视化检测海产品中的活副溶血性弧菌,该研究将叠氮溴化丙锭(propidium monoazide,PMA)染料与跨越式滚环等温扩增(saltatory rolling circle amplification,SRCA)技术相结合,以toxR为靶基因,设计并筛选引物,成功建立可视化PMA⁃SRCA检测方法,并评估所建立的方法。研究表明,12株副溶血性弧菌样品呈现阳性,29株非副溶血性弧菌样品呈现阴性,表明该方法特异性良好。该方法灵敏度为3.2×100 CFU/mL,高于可视化PMA⁃环介导等温扩增(3.2×101 CFU/mL)和可视化PMA⁃聚合酶链式反应(3.2×102 CFU/mL)的灵敏度。对76份市售海鲜样品进行检测,该方法的敏感性为100.00%,特异性为92.00%,符合率为97.37%。综上所述,可视化PMA⁃SRCA检测方法可检测活的副溶血性弧菌,具有良好的特异性、较优异的灵敏度。To visually detect viable Vibrio parahaemolyticus in seafood,this study employed the propidium monoazide(PMA)dye in conjunction with saltatory rolling circle amplification(SRCA)technology.The prim⁃ers were designed and screened targeting the toxR gene.The PMA⁃SRCA visual detection method was success⁃fully established and evaluated.The results showed that 12 Vibrio parahaemolyticus samples were positive and 29 non⁃Vibrio parahaemolyticus samples were negative,indicating that the method has good specificity.More⁃over,this method displayed a sensitivity of 3.2×100 CFU/mL,higher than that of both visual PMA⁃LAMP(3.2×101 CFU/mL)and visual PMA⁃PCR(3.2×102 CFU/mL).In the detection of 76 seafood samples in total,this method achieved a sensitivity,specificity,and coincidence rate of 100.00%,92.00%,and 97.37%,respec⁃tively.In conclusion,the PMA⁃SRCA visual detection method exhibits exceptional specificity and remarkable sensitivity in detecting viable Vibrio parahaemolyticus.

关 键 词：跨越式滚环等温扩增 副溶血性弧菌 叠氮溴化丙锭 可视化 检测 

分 类 号：TS254.7[轻工技术与工程—水产品加工及贮藏工程]