Molecular detection of hepatitis A virus in urban sewage in Rio de Janeiro, Brazil
Molecular detection of hepatitis A virus in urban sewage in Rio de Janeiro, Brazil
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DOI：
10.1111/j.1472-765x.2007.02164.x
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发表时间：
2007-08-01
影响因子：
2.4
通讯作者：
Gaspar, A. M. C.
中科院分区：
文献类型：
--
作者：
Villar, L. M.;de Paula, V. S.;Gaspar, A. M. C.
目的：进行了为期一年的调查，以检查甲型肝炎病毒（HAV）的患病率，基因型的分布及其与细菌指标的关系，在原材料和处理后的污水samples.Methods和Results：50个污水样本（原材料= 25和处理= 25），每月两次从一个污水处理厂在里约热内卢。通过吸附至负电性膜然后超滤进行病毒浓缩。采用巢式逆转录-聚合酶链反应（RT-PCR）和实时荧光定量PCR检测病毒RNA，阳性产物直接测序。还测定了总大肠菌群和粪便大肠菌群浓度。巢式RT-PCR检测HAV RNA阳性率为32%（16/50），其中16%（8/50）的HAV RNA来自污水处理厂。实时荧光定量PCR检测结果显示，50份样品中有46份（92%）检出HAV RNA，其中24份来自处理后的污水。系统发育分析将9个分离株（56%）归类为IA亚型，7个分离株（44%）归类为IB亚型。结论：Real-time PCR比巢式RT-PCR更敏感;描述了IA和IB亚型的存在，细菌指标不能用于预测污水中HAV的存在。这些结果表明，HAV仍然存在于污水处理后的环境中，并可能在维持HAV感染的地方性中起重要作用。
Aims: A one-year survey was conducted to examine hepatitis A virus (HAV) prevalence, distribution of genotypes and their relationship to bacterial indicators in raw and treated sewage samples.Methods and Results: Fifty sewage samples (raw = 25 and treated = 25) were collected twice monthly from one sewage treatment plant in Rio de Janeiro. Virus concentration was performed by adsorption to an electronegative membrane followed by ultrafiltration. Viral RNA was detected by nested reverse transcriptase-polymerase chain reaction (RT-PCR) and real-time PCR and positive products were directly sequenced. Total and faecal coliform concentrations were also determined. By nested RT-PCR, HAV RNA was detected in 16/50 (32%) and eight (16%) of them were found in treated sewage samples. By real-time PCR, HAV RNA was detected in 46/50 (92%) samples and 24 were from treated sewage. Phylogenetic analyses classified nine isolates (56%) as subgenotype IA and seven (44%) as IB.Conclusions: Real-time PCR was more sensitive than nested RT-PCR; the presence of subgenotypes IA and IB was described and bacterial indicators cannot be used to predict HAV presence in sewage.Significance and Impact of the Study: These results demonstrated that HAV still remains in the environment after sewage treatment and could play an important role in maintaining the endemicity of HAV infection.