Jiménez-Clavero Miguel Angel, Agüero Montserrat, Rojo Gema, Gómez-Tejedor Concepción
Departamento de Enfermedades Emergentes, Laboratorio Central de Veterinaria, Ctra. Algete, km 8, 28110, Algete (Madrid), Spain.
J Vet Diagn Invest. 2006 Sep;18(5):459-62. doi: 10.1177/104063870601800505.
West Nile virus represents an emerging threat for animal and human health worldwide. This virus exhibits a marked genetic variation, with at least 2 distinct evolutionary lineages. Lineage 1 has been recognized in Africa, Asia, Europe, Oceania, and more recently in the Americas, whereas lineage 2 is restricted to Africa. Perhaps for this reason, the available real-time RT-PCR methods for detecting West Nile virus genome have mainly focused on lineage 1. However, both viruses may potentially be spread beyond their endemic areas by migratory birds. This report describes a new real-time reverse transcription-PCR (RT-PCR) method based on a 5'-Taq nuclease-3' minor groove binder DNA probe (TaqMan MGB) that allows the detection of a wide range of West Nile virus isolates, including both lineages 1 and 2. This method was able to detect West Nile viruses from different origins (North and Central Africa, Middle East, Europe, and North America), whereas other flaviviruses (Usutu, Dengue, Yellow fever) analyzed in parallel remained negative. The sensitivity achieved by this assay was 10(-2)-10(-3) pfu/tube. This method, which can be performed in 96-well format, could be suitable for the large-scale surveillance of West Nile virus in areas where both lineages can potentially spread.
西尼罗河病毒对全球动物和人类健康构成了新出现的威胁。这种病毒表现出明显的基因变异，至少有2个不同的进化谱系。谱系1在非洲、亚洲、欧洲、大洋洲以及最近在美洲均有发现，而谱系2仅限于非洲。也许正因如此，现有的用于检测西尼罗河病毒基因组的实时逆转录聚合酶链反应（RT-PCR）方法主要集中在谱系1。然而，这两种病毒都有可能通过候鸟传播到其流行地区之外。本报告描述了一种基于5'-Taq核酸酶-3'小沟结合剂DNA探针（TaqMan MGB）的新型实时逆转录PCR（RT-PCR）方法，该方法能够检测包括谱系1和谱系2在内的多种西尼罗河病毒分离株。该方法能够检测来自不同来源（北非、中非、中东、欧洲和北美）的西尼罗河病毒，而同时分析的其他黄病毒（乌苏图病毒、登革病毒、黄热病毒）检测结果均为阴性。该检测方法的灵敏度为10(-2)-10(-3) 空斑形成单位/管。这种可在96孔板中进行的方法，可能适用于两个谱系都有可能传播的地区对西尼罗河病毒进行大规模监测。
