水产品和水体中灿烂弧菌现场可视化环介导等温扩增快检方法的建立及应用  

Establishment and Application of Visual LAMP Methods for Quick Detection of Vibrio Splendidus in Aquatic Products and Water

作　　者：田卓 郑秋月 麻丽丹 郑文杰 尚德静[1] 曹际娟 TIAN Zhuo;ZHENG Qiu-Yue;MA Li-Dan;ZHENG Wen-Jie;SHANG De-Jing;CAO Ji-Juan(Cell Biology Major,College of Life Sciences,Liaoning Normal University,Dalian 116033,China;Key Laboratory of Biotechnology and Resource Utilization of Ministry of Education,Dalian University for Nationalities,Dalian 116600,Liaoning,China;Dalian Customs Technology Center,Dalian 116000,Liaoning,China;Dandong Customs Comprehensive Technical Service Center,Dandong 118000,Liaoning,China;Biology Major,College of Life Sciences,Tianjin Normal University,Tianjin 300387,China)

机构地区：[1]辽宁师范大学生命科学学院细胞生物学专业,辽宁大连116033 [2]大连民族大学生物技术与资源利用教育部重点实验室,辽宁大连116600 [3]大连海关技术中心,辽宁大连116000 [4]丹东海关综合技术服务中心,辽宁丹东118000 [5]天津师范大学生命科学学院生物学专业,天津300387

出　　处：《中国生物化学与分子生物学报》2022年第1期108-119,共12页Chinese Journal of Biochemistry and Molecular Biology

基　　金：辽宁省“兴辽英才计划”(No.XLYC2002106);大连市高层次人才创新支持计划(No.2019CT09)资助*。

摘　　要：基于环介导等温扩增(LAMP)技术建立水产品和养殖水域中灿烂弧菌现场可视化的快速、简便检测方法。以灿烂弧菌等作为研究对象,以灿烂弧菌的toxR基因作为靶基因,确定煮沸法为适合于弧菌基因组DNA提取的快捷方法,优化筛选的引物可以特异地检测灿烂弧菌,检测核酸浓度的灵敏度可以达到10^(-9) g/L,并且结果稳定、可靠。采用该方法进行实际样品的检测应用,在558份水体样品和655份水产品样品中,灿烂弧菌阳性检出率分别为0.54%和0.46%,与实时荧光PCR方法的检测结果符合率为100%。低技术要求,低设备成本以及短检测时间使得可视化LAMP快检方法成为现场可使用的一个理想选择,这对于水产养殖业来说也更方便。A visualized,rapid,simple method was developed based on loop-mediated isothermal amplification(LAMP)to detect Vibrio splendidus in aquatic products and aquaculture waters.Taking Vibrio splendidus as the research object,and the toxR gene of Vibrio splendidus as the target gene,the boiling method was determined as a fast method suitable for the genomic DNA extraction of Vibrio splendidus.The optimized and screened primers could specifically detect Vibrio splendidus,and the sensitivity of nucleic acid concentration detection could reach 10^(-9)g/L.We show that the results were stable and reliable.In 558 water samples and 655 aquatic product samples,the positive detection rate of Vibrio brilliant was 0.54%and 0.46%,respectively,and the coincidence rate with the real-time PCR method was 100%.Low technical requirements,low equipment costs,and short detection time render the visual LAMP rapid detection method an ideal choice in field diagnosis and convenience for the aquaculture industry.

关 键 词：灿烂弧菌 水产品 水体 可视化 环介导等温扩增 

分 类 号：Q93-331[生物学—微生物学]