Rotavirus A (RVA) is a diarrheal pathogen affecting children under age five, particularly in developing and underdeveloped regions of the world due to malnutrition, poor healthcare and hygienic conditions. Water and food contamination are found to be major sources of diarrheal outbreaks. Pakistan is one of the countries with high RVA related diarrhea burden but with insufficient surveillance system. The aim of this study was to gauge the RVA contamination of major open sewerage collecting streams and household water supplies in two major metropolitan cities of Pakistan. Three concentration methods were compared using RNA purity and concentration as parameters, and detection efficiency of the selected method was estimated. Water samples were collected from 21 sites in Islamabad and Rawalpindi in two phases during the year 2014-2015. Meteorological conditions were recorded for each sampling day and site from Pakistan Meteorological Department (PMD). Nested PCR was used to detect the presence of RVA in samples targeting the VP7 gene. Logistic regression was applied to assess the association of weather conditions with RVA persistence in water bodies. Statistical analysis hinted at a temporal and seasonal pattern of RVA detection in water. Phylogenetic analysis of selected isolates showed a close association of environmental strains with clinical RVA isolates from hospitalized children with acute diarrhea during the same period. This is the first scientific report cataloging the circulating RVA strains in environmental samples from the region. The study highlights the hazards of releasing untreated sewerage containing potentially infectious viral particles into collecting streams, which could become a reservoir of multiple pathogens and a risk to exposed communities. Moreover, routine testing of these water bodies can present an effective surveillance system of circulating viral strains in the population.

译文

轮状病毒A(RVA)是一种腹泻病原体,由于营养不良,医疗保健和卫生状况不佳,尤其在世界的发展中和欠发达地区影响五岁以下的儿童。发现水和食物污染是腹泻暴发的主要来源。巴基斯坦是RVA相关的腹泻负担高但监测系统不足的国家之一。这项研究的目的是评估巴基斯坦两个主要大城市的主要露天排污收集流和家庭供水的RVA污染。以RNA纯度和浓度为参数,比较了三种浓缩方法,并评估了所选方法的检测效率。 2014-2015年分两个阶段从伊斯兰堡和拉瓦尔品第的21个地点收集了水样。巴基斯坦气象局(PMD)记录了每个采样日和每个站点的气象条件。巢式PCR用于检测靶向VP7基因的样品中RVA的存在。应用逻辑回归分析评估天气状况与水体中RVA持久性之间的关系。统计分析提示了水中RVA检测的时间和季节模式。对选定菌株的系统进化分析表明,同期来自急性腹泻住院儿童的环境菌株与临床RVA菌株密切相关。这是第一份分类该地区环境样品中循环中的RVA菌株的科学报告。这项研究强调了将未经处理的含有潜在感染性病毒颗粒的污水释放到收集流中的危害,这可能成为多种病原体的储存库,并给裸露的社区带来风险。此外,对这些水体的常规检测可以提供一种有效的人群中循环病毒株监测系统。

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