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应用细胞培养和聚合酶链反应检测科特迪瓦约普贡水样中的肠道病毒。

Detection of Enteroviruses in Water Samples from Yopougon, Côte d'Ivoire by Cell Culture and Polymerase Chain Reaction.

机构信息

Unité des virus du système nerveux, Département virus épidémique, Institut Pasteur de Cote d'Ivoire, BP 490, Abidjan 01, Ivory Coast.

UFR sciences et gestion de l'environnement, Laboratoire des sciences de l'environnement université Nangui Abrogoua, BP 801, Abidjan 02, Ivory Coast.

出版信息

Food Environ Virol. 2014 Mar;6(1):23-30. doi: 10.1007/s12560-013-9130-4. Epub 2013 Nov 2.

Abstract

The objective of this study was to compare sensitivities of enterovirus isolation from wastewater in different cell lines as well as to compare the sensitivity and specificity of isolation in cell culture with direct detection by reverse transcription polymerase chain reaction (RT-PCR). Sixty-eight samples of wastewaters were collected between September 2008 and January 2009 in Yopougon, Abidjan. Enteroviruses were concentrated according to World Health Organization recommendations. Viruses were inoculated into various cell lines while direct RT-PCR was performed on water concentrates. The buffalo green monkey kidney cell line was the most sensitive with 58.8 % of viral isolation. This was followed by the rhabdomyosarcoma cell line with sensitivity of 51.6 %, with human epidermoid carcinoma cell line showing sensitivity of 50 % and fibroblastic cells derived from transgenic mice LTK-1 (L20B) cell showing 23.50 % sensitivity. However, a lower specificity of 2.9 % was observed with the L20B cell line. 44.1 % of the samples were positive by direct RT-PCR detection while 51.47 % samples were positive by using RT-PCR on infected cell cultures. No difference in percentage positivity was observed using RT-PCR on infected tissue culture isolates or using RT-PCR directly on wastewater samples.

摘要

本研究的目的是比较在不同细胞系中从废水中分离肠道病毒的敏感性,以及比较细胞培养物中的分离的敏感性和特异性与逆转录聚合酶链反应(RT-PCR)直接检测的敏感性和特异性。2008 年 9 月至 2009 年 1 月期间,在阿比让的约普贡收集了 68 份废水样本。根据世界卫生组织的建议,对肠道病毒进行浓缩。将病毒接种到各种细胞系中,同时对水样进行直接 RT-PCR。水牛绿猴肾细胞系的病毒分离敏感性最高,为 58.8%。紧随其后的是横纹肌肉瘤细胞系,敏感性为 51.6%,人表皮样癌细胞系敏感性为 50%,来自转基因小鼠 LTK-1(L20B)的成纤维细胞敏感性为 23.50%。然而,L20B 细胞系的特异性较低,为 2.9%。直接 RT-PCR 检测有 44.1%的样本呈阳性,而用感染细胞培养物的 RT-PCR 检测有 51.47%的样本呈阳性。使用感染组织培养物分离物的 RT-PCR 或直接使用 RT-PCR 检测废水样本,阳性百分比无差异。

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