Ferreira F F M, Guimarães F R, Fumian T M, Victoria M, Vieira C B, Luz S, Shubo T, Leite J P G, Miagostovich M P
Laboratório de Virologia Comparada e Ambiental, Instituto Oswaldo Cruz, Rio de Janeiro, Pavilhão Hélio & Peggy Pereira, Fundação Oswaldo Cruz, Avenida Brasil, 4365-Maguinhos 21040-360, Rio de Janeiro RJ, Brazil.
Water Sci Technol. 2009;60(3):633-42. doi: 10.2166/wst.2009.413.
Rotaviruses A (RV-A) infection is the most common cause of acute diarrheal diseases in infants and the dissemination of these viruses in the environment represents a public health hazard. The present study aims to evaluate reverse transcription-polymerase chain reaction (RT-PCR) based protocols for the detection of RV-A genes in different types of environmental samples. RV-A were concentrated by the adsorption-elution method using negatively charged membranes associated with a Centriprep Concentrator 50. The RV-A VP4, VP7 and VP6 genes were detected using RT-PCR in river water from the Amazon Hydrographic basin (Northern region) and from wastewater in a sewage treatment plant in Rio de Janeiro (Southeast region), Brazil. RV-A were successfully detected in water environmental samples by the methods used. The detection of the VP6 gene by RT-PCR was the most sensitive for detecting RV-A in environmental samples (44.0%), when compared to the detection of the VP4 (33.3%) and VP7 (25.3%) genes. Based on nucleotide sequence and phylogenetic analysis of the partial VP6 gene, 22 environmental samples were determined to be subgroup II (Wa-like). These results indicate that analysis of environmental samples could possibly make a valuable contribution to studies on the epidemiology of RV-A.
A组轮状病毒（RV-A）感染是婴儿急性腹泻疾病的最常见病因，这些病毒在环境中的传播构成公共卫生危害。本研究旨在评估基于逆转录聚合酶链反应（RT-PCR）的方案，用于检测不同类型环境样本中的RV-A基因。使用与Centriprep Concentrator 50相关的带负电荷膜，通过吸附-洗脱法浓缩RV-A。使用RT-PCR在巴西亚马逊水文流域（北部地区）的河水和里约热内卢（东南部地区）一家污水处理厂的废水中检测RV-A的VP4、VP7和VP6基因。通过所使用的方法在水环境样本中成功检测到RV-A。与VP4基因（33.3%）和VP7基因（25.3%）的检测相比，RT-PCR检测VP6基因对检测环境样本中的RV-A最为敏感（44.0%）。基于部分VP6基因的核苷酸序列和系统发育分析，确定22个环境样本为II亚组（Wa样）。这些结果表明，环境样本分析可能对RV-A流行病学研究做出有价值的贡献。
