Tong Qun-Bo, Chen Rui, Zhang Yi, Yang Guo-Jing, Kumagai Takashi, Furushima-Shimogawara Rieko, Lou Di, Yang Kun, Wen Li-Yong, Lu Shao-Hong, Ohta Nobuo, Zhou Xiao-Nong
Institute of Parasitic Diseases, Zhejiang Academy of Medical Sciences, Hangzhou, P.R. China.
National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention, Shanghai, P.R. China; Key Laboratory for Parasite and Vector Biology, MOH, P.R. China.
Acta Trop. 2015 Jan;141(Pt B):170-7. doi: 10.1016/j.actatropica.2014.01.006. Epub 2014 Feb 2.
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 28S 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.
尽管血吸虫病在全球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防治方面已取得显著成就。该病在12个流行省份中的5个已被消除,其余流行地区的患病率非常低且正朝着消除的方向发展。迫切需要一种快速、灵敏的方法来监测感染性钉螺的分布情况。我们应用了一种针对28S rDNA的环介导等温扩增(LAMP)检测法,用于快速、有效地检测感染和处于潜伏期感染的日本血吸虫DNA在钉螺中的情况。LAMP方法的检测限为100 fg日本血吸虫基因组DNA。为了促进该方法在现场的应用,LAMP检测法被用于检测野外采集的钉螺混合样本中的感染情况。在混合样本检测中,从28个流行地区收集钉螺,根据最大混合样本量估计,每个地区收集50只钉螺混合在一起,共同碾碎,并从每个混合样本中提取DNA作为LAMP检测的模板。根据混合样本检测公式,LAMP检测法检测到的西马村阳性混合样本比例和感染钉螺的阳性比例分别达到66.67%和1.33%,而黑泥、洪家、阳江和黄山村的分别为33.33%和0.67%,团洲和苏辽村的分别为16.67%和0.33%。其余21个监测现场均为阴性结果。基于LAMP检测法检测到的感染日本血吸虫的钉螺阳性比例,使用ArcMap构建了血吸虫病传播风险图,这将为高风险地区的监测和应对策略提供指导。