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对诊断为急性弛缓性麻痹的儿童粪便标本中肠道病毒检测算法的比较。

Comparison of Algorithms for the Detection of Enteroviruses in Stool Specimens from Children Diagnosed with Acute Flaccid Paralysis.

作者信息

Adeniji J A, Ayeni F A, Ibrahim A, Tijani K A, Faleye T O C, Adewumi M O

机构信息

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

WHO National Polio Laboratory, University of Ibadan, Ibadan, Oyo State, Nigeria.

出版信息

J Pathog. 2017;2017:9256056. doi: 10.1155/2017/9256056. Epub 2017 Dec 28.

Abstract

This study was designed to compare both the cell culture dependent and independent enterovirus detection algorithms recommended by the WHO and assess how either might impact our perception of the diversity of enterovirus types present in a sample. Sixteen paired samples (16 isolates from RD cell culture and their corresponding stool suspension, i.e., 32 samples) from AFP cases in Nigeria were analyzed in this study. All the samples were subjected to RNA extraction, cDNA synthesis, the WHO recommended RT-snPCR, and its modification. Amplicons were sequenced and strains identified. Enterovirus diversity was the same between the isolates and fecal suspension for the control and five of the samples. It was, however, different for the remaining 10 (62.5%) samples. Nine (CV-B4, E6, E7, E13, E14, E19, E29, EV-B75, and EV-B77) and five (CV-A1, CV-A11, CV-A13, EV-C99, and PV2) EV-B and EV-C types, respectively, were detected. Particularly, E19 and EV-B75 were only recovered from the isolates while E14, EV-B77, CV-A11, and CV-A13 were only recovered from fecal suspension. Both the cell culture dependent and independent protocols bias our perception of the diversity of enterovirus types present in a sample. Hence, effort should be directed at harmonizing both for increased sensitivity.

摘要

本研究旨在比较世界卫生组织推荐的依赖细胞培养和不依赖细胞培养的肠道病毒检测算法,并评估这两种算法如何影响我们对样本中存在的肠道病毒类型多样性的认知。本研究分析了来自尼日利亚急性弛缓性麻痹(AFP)病例的16对样本(从RD细胞培养中分离出的16株病毒及其相应的粪便悬液,即32个样本)。所有样本均进行RNA提取、cDNA合成、世界卫生组织推荐的逆转录巢式聚合酶链反应(RT-snPCR)及其改良方法。对扩增子进行测序并鉴定毒株。对照样本和5个样本的病毒分离株与粪便悬液中的肠道病毒多样性相同。然而,其余10个样本(62.5%)的情况则不同。分别检测到9种B组肠道病毒(CV-B4、E6、E7、E13、E14、E19、E29、EV-B75和EV-B77)和5种C组肠道病毒(CV-A1、CV-A11、CV-A13、EV-C99和PV2)。特别值得注意的是,E19和EV-B75仅从病毒分离株中检出,而E14、EV-B77、CV-A11和CV-A13仅从粪便悬液中检出。依赖细胞培养和不依赖细胞培养的方案都会使我们对样本中存在的肠道病毒类型多样性的认知产生偏差。因此,应致力于协调这两种方法以提高检测灵敏度。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/96e2/5763071/5a1b9119b06d/JPATH2017-9256056.001.jpg

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