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,医疗保健和卫生条件差,影响五岁以下儿童,特别是在世界发展中国家和欠发达地区。发现水和食物污染是腹泻暴发的主要来源。巴基斯坦是与RVA相关的腹泻负担较高但监测系统不足的国家之一。这项研究的目的是衡量巴基斯坦两个主要大都市的主要露天排污系统收集溪流和家庭供水的RVA污染。以RNA纯度和浓度为参数,比较了三种浓度方法,并估算了所选方法的检测效率。在2014-2015年期间,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。记录了巴基斯坦气象局 (PMD) 每个采样日和地点的气象条件。巢式PCR用于检测靶向VP7基因的样品中RVA的存在。应用逻辑回归来评估天气条件与水体中RVA持久性的关联。统计分析暗示了水中RVA检测的时间和季节模式。对选定分离株的系统发育分析表明,环境菌株与同期住院急性腹泻儿童的临床RVA分离株密切相关。这是第一份对该地区环境样品中循环的RVA菌株进行分类的科学报告。该研究强调了将含有潜在传染性病毒颗粒的未经处理的污水排放到收集流中的危害,这可能成为多种病原体的储存库,并对暴露的社区构成风险。此外,对这些水体的常规测试可以为人群中循环病毒株提供有效的监视系统。

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