Wijegunawardana Asha Dilrukshi, Gunawardane Nilmini Silva, Hapuarachchi Chanditha, Manamperi Aresha, Gunawardena Kithsiri, Abeyewickrama Wimaladharma, Latif Baha
Molecular Medicine Unit, Faculty of Medicine, University of Kelaniya, Ragama, Sri Lanka.
Asian Pac J Trop Biomed. 2013 May;3(5):381-7. doi: 10.1016/S2221-1691(13)60080-5.
To compare Wuchereria bancrofti (W. bancrofti) infection rates of Culex quinquefasciatus, using dissection and PCR-ELISA in two consecutive time periods (from 2007 to 2008 and from 2008 to 2009).
Mosquitoes were collected in 30 sentinel and 15 non-sentinel sites in 15 Medical Officer of Health areas of Gampaha District known for the presence of W. bancrofti transmission in two consecutive time period of 2007 to 2008 and 2008 to 2009. Captured mosquitoes were dissected to determine the W. bancrofti larvae (L1, L2, L3). PCR was carried out using DNA extracted from mosquito pools (15 body parts/pool) utilizing the primers specific for Wb-SspI repeat. PCR products were analyzed by hybridization ELISA using fluorescein-labeled wild type specific probes. The prevalence of infected/infective mosquitoes in PCR pools (3 pools/site) was estimated using the PoolScreen™ algorithm and a novel probability-based method.
Of 45 batches of mosquitoes dissected, W. bancrofti infected mosquitoes were found in 19 and 13 batches, with an infection rate of 13.29% and 3.10% with mean larval density of 8.7 and 1.0 larvae per mosquito for two study periods in the Gampaha District. Total of 405 pools of head, thorax and abdomen were processed by PCR-ELISA for each year. Of these, 51 and 31 pools were positive for W. bancrofti in the two study periods respectively. The association of dissection based prevalence rates with PCR based rates as determined by the Pearson correlation coefficient were 0.176 and 0.890 respectively for the two periods.
Data indicate that PCR-ELISA is more sensitive than the traditional dissection techniques for monitoring transmission intensity.
比较在两个连续时间段（2007年至2008年以及2008年至2009年）使用解剖法和聚合酶链反应 - 酶联免疫吸附测定法（PCR - ELISA）检测致倦库蚊中班氏吴策线虫（W. bancrofti）的感染率。
在甘帕哈区15个卫生医疗区域的30个监测点和15个非监测点收集蚊子，这些区域已知存在班氏吴策线虫传播，时间为连续的两个时间段，即2007年至2008年以及2008年至2009年。捕获的蚊子进行解剖以确定班氏吴策线虫幼虫（L1、L2、L3）。使用从蚊群（每群15个身体部位）提取的DNA进行PCR，所用引物针对Wb - SspI重复序列。PCR产物通过使用荧光素标记的野生型特异性探针的杂交酶联免疫吸附测定法进行分析。使用PoolScreen™算法和一种基于概率的新方法估计PCR样本池（每个地点3个样本池）中受感染/具感染性蚊子的流行率。
在解剖的45批蚊子中，在19批和13批中发现了感染班氏吴策线虫的蚊子，在甘帕哈区的两个研究时间段内，感染率分别为13.29%和3.10%，平均幼虫密度分别为每只蚊子8.7条和1.0条幼虫。每年通过PCR - ELISA处理405个头、胸和腹部样本池。其中，在两个研究时间段内分别有51个和31个样本池检测出班氏吴策线虫呈阳性。两个时间段通过Pearson相关系数确定的基于解剖的流行率与基于PCR的流行率之间的关联分别为0.176和0.890。
数据表明，对于监测传播强度，PCR - ELISA比传统解剖技术更敏感。
