Emerging Pathogens Laboratory, Fondation Mérieux, 21 Avenue Tony Garnier, 69007 Lyon, France.
J Virol Methods. 2011 Sep;176(1-2):74-7. doi: 10.1016/j.jviromet.2011.06.003. Epub 2011 Jun 15.
A one-step real time quantitative RT-PCR (qRT-PCR) assay was developed to detect all published Dugbe virus (DUGV) genomes of the Nairovirus genus. Primers and probes were designed to detect specific sequences on the most conserved regions of the S segment. The limit of detection of the assay was 10 copies per reaction which is an improvement of 3 log(10)FFU/mL over the sensitivity of conventional RT-PCR. The specificity of the primers and probe was confirmed with the closely related Nairoviruses CCHFV and Hazara virus, and on the non-related viruses Coronavirus and Influenza A virus. This qRT-PCR assay was used to screen nucleic acids extracted from 498 ticks collected in the Republic of Chad. One sample was found positive suggesting that DUGV is present in this part of the world. The molecular assay developed in this study is sensitive, specific and rapid and can be used for research and epidemiological studies.
建立了一种一步法实时定量 RT-PCR(qRT-PCR)检测方法，用于检测纳罗病毒属中所有已发表的杜格贝病毒(DUGV)基因组。设计了引物和探针，以检测 S 片段上最保守区域的特定序列。该检测方法的检测限为每个反应 10 个拷贝，比传统 RT-PCR 的灵敏度提高了 3 个对数(10)FFU/mL。用密切相关的纳罗病毒 CCHFV 和 Hazara 病毒以及不相关的冠状病毒和甲型流感病毒验证了引物和探针的特异性。该 qRT-PCR 检测方法用于筛选在乍得共和国采集的 498 只蜱中的核酸。发现一个样本呈阳性，提示 DUGV 存在于世界的这一地区。本研究中开发的分子检测方法具有敏感性、特异性和快速性，可用于研究和流行病学研究。
