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目的探讨环介导等温扩增技术(LAMP)的特异性与灵敏度,并评价其在在霍乱监测中的应用价值。方法 2012—2013年每月从珠江海珠区沿岸6个河水监测点、6个河涌及养殖场污水监测点、2个市场海水产品监测点采集珠江水、污水及市售海水产品等外环境霍乱监测样本,利用LAMP技术进行检测,以传统分离培养法作对照,并对LAMP技术进行灵敏度及特异度试验。结果共采集403份样本,荧光LAMP法共检出8份霍乱孤菌阳性,分离培养法检出6份霍乱孤菌阳性,经统计学分析,LAMP法和传统培养法检测阳性率的差异无统计学意义(P>0.05);荧光LAMP法灵敏度为100.00%,特异度为99.49%;LAMP法对霍乱弧菌的检测限可达2.88×102CFU/mL,符合率为99.50%。结论 LAMP技术对霍乱弧菌检测的特异性强、灵敏度高,快速简便,对基层疾控部门及临床实验室霍乱疫情的监控有实际意义。
Objective To investigate the specificity and sensitivity of loop-mediated isothermal amplification (LAMP) and to evaluate its value in monitoring cholera. Methods From 2012 to 2013, cholera was collected from 6 river water monitoring points, 6 river surges and farm sewage monitoring points along the coast of Haizhu District, Pearl River and monitoring points of two market marine products to collect external environmental cholera such as pearl river water, sewage water and commercial seawater products Monitoring samples, the use of LAMP technology for testing, the traditional separation culture as a control, and LAMP technology for sensitivity and specificity tests. Results A total of 403 samples were collected. Eight cholera spp. Positive were detected by fluorescence LAMP method and six strains of V. cholerae were detected by isolation culture method. There was no statistic difference between the positive rate of LAMP assay and traditional culture method (P> 0.05). The sensitivity of fluorescence LAMP method was 100.00% and the specificity was 99.49%. The detection limit of LAMP method for Vibrio cholerae reached 2.88 × 102CFU / mL with the coincidence rate of 99.50%. Conclusion The LAMP technique has strong specificity, high sensitivity and quickness and convenience for the detection of Vibrio cholerae, and has practical significance for the monitoring of cholera epidemics in grassroots disease control departments and clinical laboratories.