A new surveillance and response tool: Risk map of infected Oncomelania hupensis detected by Loop-mediated isothermal amplification (LAMP) from pooled samples
A new surveillance and response tool: Risk map of infected Oncomelania hupensis detected by Loop-mediated isothermal amplification (LAMP) from pooled samples
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DOI：
10.1016/j.actatropica.2014.01.006
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发表时间：
2015-01-01
期刊：
影响因子：
2.7
通讯作者：
Zhou, Xiao-Nong
中科院分区：
文献类型：
--
作者：
Tong, Qun-Bo;Chen, Rui;Zhou, Xiao-Nong
虽然血吸虫病在世界范围内仍然是一个严重的健康问题，但中华人民共和国在血吸虫病控制方面取得了重大成就。在12个流行省份中，已有5个省消灭了该病，其余流行地区的流行率很低，正朝着消灭的方向发展。迫切需要一种快速、灵敏的方法来监测感染的甲型钉螺的分布。采用285 rDNA为靶点的环介导等温扩增（LAMP）方法，快速有效地检测感染和预感染钉螺中的日本血吸虫DNA。LAMP方法检测限为100 fg的日本血吸虫基因组DNA。为了推广该方法在田间的应用，采用LAMP法对田间采集的钉螺进行了感染检测。在混合样本检测中，从28个流行区收集钉螺，根据最大池大小估计每个区收集50只钉螺，粉碎在一起，从每个混合样本中提取DNA作为LAMP检测模板。根据合并样本检测公式，西马村合并样本阳性率和LAMP检测阳性率分别为66.67%和1.33%，黑尼村、洪家村、阳江村和黄山村分别为33.33%和0.67%，团州村和苏寮村分别为16.67%和0.33%。其余21个监测场址的结果为阴性。利用ArcMap软件，根据LAMP法检测到的血吸虫感染阳性比例，构建血吸虫病传播风险图，为血吸虫病高风险地区的监测和应对策略提供指导。(C) 2014 Elsevier B.V.版权所有
Although schistosomiasis remains a serious health problem worldwide, significant achievements in schistosomiasis control has been made in the People's Republic of China. The disease has been eliminated in five out of 12 endemic provinces, and the prevalence in remaining endemic areas is very low and is heading toward elimination. A rapid and sensitive method for monitoring the distribution of infected Oncomelania hupensis is urgently required. We applied a loop-mediated isothermal amplification (LAMP) assay targeting 285 rDNA for the rapid and effective detection of Schistosoma japonicum DNA in infected and prepatent infected O. hupensis snails. The detection limit of the LAMP method was 100 fg of S. japonicum genomic DNA. To promote the application of the approach in the field, the LAMP assay was used to detect infection in pooled samples of field-collected snails. In the pooled sample detection, snails were collected from 28 endemic areas, and 50 snails from each area were pooled based on the maximum pool size estimation, crushed together and DNA was extracted from each pooled sample as template for the LAMP assay. Based on the formula for detection from pooled samples, the proportion of positive pooled samples and the positive proportion of O. hupensis detected by LAMP of Xima village reached 66.67% and 1.33%, while those of Heini, Hongjia, Yangjiang and Huangshan villages were 33.33% and 0.67%, and those of Tuanzhou and Suliao villages were 16.67% and 0.33%, respectively. The remaining 21 monitoring field sites gave negative results. A risk map for the transmission of schistosomiasis was constructed using ArcMap, based on the positive proportion of O. hupensis infected with S. japonicum, as detected by the LAMP assay, which will form a guide for surveillance and response strategies in high risk areas. (C) 2014 Elsevier B.V. All rights reserved.