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尼日利亚急性弛缓性麻痹病例粪便标本中 RD-L20B 细胞培养阴性样本中肠道病毒 C 大量存在,且具有地理定义性。

Abundance of enterovirus C in RD-L20B cell culture-negative stool samples from acute flaccid paralysis cases in Nigeria is geographically defined.

机构信息

Department of Medical Microbiology and Parasitology, Obafemi Awolowo University, Ile-Ife, Osun State, Nigeria.

Department of Virology, College of Medicine, University of Ibadan, Ibadan, Oyo State, Nigeria.

出版信息

J Med Microbiol. 2018 Jun;67(6):854-865. doi: 10.1099/jmm.0.000737. Epub 2018 Apr 30.

Abstract

PURPOSE

We recently showed that enteroviruses (EVs) andenterovirus species C (EV-C) in particular were abundant in faecal samples from children who had been diagnosed with acute flaccid paralysis (AFP) in Nigeria but declared to be EV-free by the RD-L20B cell culture-based algorithm. In this study, we investigated whether this observed preponderance of EVs (and EV-Cs) in such samples varies by geographical region.

METHODOLOGY

One hundred and eight samples (i.e. 54 paired stool suspensions from 54 AFP cases) that had previously been confirmed to be negative for EVs by the WHO-recommended RD-L20B cell culture-based algorithm were analysed. The 108 samples were made into 54 pools (27 each from North-West and South-South Nigeria). All were subjected to RNA extraction, cDNA synthesis and the WHO-recommended semi-nested PCR assay and its modifications. All of the amplicons were sequenced, and the enteroviruses identified, using the enterovirus genotyping tool and phylogenetic analysis.

RESULTS

EVs were detected in 16 (29.63 %) of the 54 samples that were screened and successfully identified in 14 (25.93 %). Of these, 10 were from North-West and 4 were from South-South Nigeria. One (7.14 %), 2 (14.29 %) and 11 (78.57 %) of the strains detected were EV-A, EV-B and EV-C, respectively. The 10 strains from North-West Nigeria included 7 EV types, namely CV-A10, E29, CV-A13, CV-A17, CV-A19, CV-A24 and EV-C99. The four EV types recovered from South-South Nigeria were E31, CV-A1, EV-C99 and EV-C116.

CONCLUSION

The results of this study showed that the presence of EVs and consequently EV-Cs in AFP samples declared to be EV-free by the RD-L20B cell culture-based algorithm varies by geographical region in Nigeria.

摘要

目的

我们最近发现,在尼日利亚被诊断为急性弛缓性麻痹(AFP)但通过 RD-L20B 细胞培养为基础的算法被宣布为无 EV 的粪便样本中,肠道病毒(EVs)和肠道病毒 C 型(EV-C)特别丰富。在这项研究中,我们研究了在这些样本中观察到的 EV(和 EV-C)的优势是否因地理位置的不同而有所不同。

方法

对 108 份先前通过世界卫生组织推荐的 RD-L20B 细胞培养为基础的算法确认的 EV 阴性样本进行了分析。这 108 个样本制成 54 个粪便悬液池(27 个来自尼日利亚的西北和南南)。所有样本均进行 RNA 提取、cDNA 合成和世界卫生组织推荐的半巢式 PCR 检测及其改良。使用肠道病毒基因分型工具和系统发育分析对所有扩增子进行测序和肠道病毒鉴定。

结果

在筛选的 54 个样本中,有 16 个(29.63%)检测到 EV,其中 14 个(25.93%)成功鉴定。其中,10 个来自尼日利亚的西北,4 个来自南南。检测到的 1 株(7.14%)、2 株(14.29%)和 11 株(78.57%)分别为 EV-A、EV-B 和 EV-C。来自尼日利亚西北的 10 株包括 7 种肠道病毒,即 CV-A10、E29、CV-A13、CV-A17、CV-A19、CV-A24 和 EV-C99。从南南尼日利亚回收的 4 种肠道病毒类型为 E31、CV-A1、EV-C99 和 EV-C116。

结论

本研究结果表明,在尼日利亚,RD-L20B 细胞培养为基础的算法宣布为无 EV 的 AFP 样本中存在 EVs,而且 EV-Cs 的存在因地理位置而异。

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