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乌拉圭的一项环境监测揭示了高度多样化的人肠道病毒 C 型和高频率的 A 型和 B 型病毒的存在。

An Environmental Surveillance in Uruguay Reveals the Presence of Highly Divergent Types of Human Enterovirus Species C and a High Frequency of Species A and B Types.

机构信息

Laboratorio de Virología Molecular, CENUR Litoral Norte, Sede Salto, Universidad de la República, Gral. Rivera 1350, 50000, Salto, Uruguay.

Laboratório de Enterovírus, Instituto Oswaldo Cruz, Fundação Oswaldo Cruz, Avda. Brasil 4365, Rio de Janeiro, 21040-360, Brazil.

出版信息

Food Environ Virol. 2018 Dec;10(4):343-352. doi: 10.1007/s12560-018-9351-7. Epub 2018 Jun 16.

Abstract

Information about Human Enterovirus circulation in Uruguay is scarce. The aim of this study was to generate the first description about their circulation in the country through the study of sewage samples collected before and after the switch from Oral Poliovirus Vaccine to Inactivated Poliovirus Vaccine. Viruses were concentrated by an adsorption-elution to a negatively charged membrane, and real-time quantitative PCR and qualitative PCR methods were used to detect, quantify, and characterize enteroviruses. Positive samples were inoculated in RD cells and two passages were performed. Additionally, RD+ samples were subsequently passed onto L20B cells. Human Enteroviruses were detected in 67.6% of the samples, with concentrations between 4.9 and 6.6 Log genomic copies per liter. 10% of positive samples replicated in RD cells, of which none in L20B cells. Molecular characterization of Human Enterovirus strains directly detected from sewage sample concentrates allowed the identification of highly divergent members of species C such as Enterovirus C99 and Coxsackievirus A13, as well as the frequent detection of species A and B members (particularly Coxsackievirus A16 and Echovirus 6, respectively). Other detected types were Coxsackievirus A2, A22, B1, B5, Echovirus 5, and 9. The characterization of viruses isolated in cell culture revealed the presence of Echovirus 6 and Coxsackievirus B3. Despite the absence of poliovirus, a wide circulation of different enterovirus types was evidenced in Uruguayan sewage samples, highlighting that the local populations are exposed to different kinds of diseases originated by several human enterovirus.

摘要

乌拉圭有关人类肠道病毒传播的信息较为匮乏。本研究旨在通过对口服脊髓灰质炎疫苗转换为灭活脊髓灰质炎疫苗前后采集的污水样本进行研究,首次对该国肠道病毒的传播情况进行描述。病毒通过吸附洗脱到带负电荷的膜上进行浓缩,采用实时定量 PCR 和定性 PCR 方法检测、定量和鉴定肠道病毒。对阳性样本进行 RD 细胞接种和两次传代,此外,RD+样本随后接种到 L20B 细胞上。在 67.6%的样本中检测到人类肠道病毒,浓度为每升 4.9 到 6.6 对数基因组拷贝。10%的阳性样本在 RD 细胞中复制,其中没有一个在 L20B 细胞中复制。从污水样本浓缩物中直接检测到的人类肠道病毒株的分子特征鉴定出了高度分化的 C 种成员,如肠道病毒 C99 和柯萨奇病毒 A13,以及 A 种和 B 种成员的频繁检测(分别为柯萨奇病毒 A16 和埃可病毒 6)。其他检测到的类型为柯萨奇病毒 A2、A22、B1、B5、埃可病毒 5 和 9。在细胞培养中分离出的病毒的特征表明存在埃可病毒 6 和柯萨奇病毒 B3。尽管没有脊髓灰质炎病毒,但在乌拉圭污水样本中发现了多种不同肠道病毒的广泛传播,这突显了当地人群面临着由多种人类肠道病毒引起的不同疾病的威胁。

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