Real-time reverse transcriptase-polymerase chain reaction quantification of West Nile virus transmitted by Culex pipiens quinquefasciatus
Real-time reverse transcriptase-polymerase chain reaction quantification of West Nile virus transmitted by Culex pipiens quinquefasciatus
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DOI：
10.4269/ajtmh.2004.71.120
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发表时间：
2004-07-01
影响因子：
3.3
通讯作者：
Higgs, S
中科院分区：
文献类型：
--
作者：
Vanlandingham, DL;Schneider, BS;Higgs, S
传播实验是媒介能力研究的重要组成部分。本研究采用实时定量逆转录聚合酶链式反应(RT-PCR)检测蚊虫口腔感染后唾液中西尼罗河病毒(West Nile Virus，WNV)的分泌量。致倦库蚊采食西尼罗河病毒感染的血液，分别于感染后第14天和第21天采集唾液。这两个时间点的病毒数量差异很大，第14天的平均当量空斑形成单位(PFU)约为30,500，第21天的平均当量空斑形成单位(PFU)约为5,800。每只蚊子分泌的病毒高达2x10(5)pfu。在这两个时间点采集的蚊子的全蚊子滴度和唾液腺的免疫荧光分析也用于补充比较。这份报告描述了第一次使用实时RT-PCR来量化蚊子唾液中的西尼罗河病毒的数量。
Transmission experiments are a critical component of vector competence studies. In this study, a real-time reverse transcriptase-polymerase chain reaction (RT-PCR) was used to enumerate the amount of West Nile virus (WNV) secreted in mosquito saliva following oral infection. Culex pipiens quinquefasciatus were allowed to feed on WNV-infected blood, and saliva was collected on days 14 and 21 post-infection (pi). The amount of virus at these two time points varied significantly, with mean equivalent plaque-forming units (pfu) of approximately 30,500 on day 14 pi and 5,800 on day 21 pi. Individual mosquitoes secreted up to 2 x 10(5) pfu of virus. Titer of whole mosquitoes and immunofluorescence assay of salivary glands from mosquitoes collected at these two time points were also used for supplemental comparison. This report describes the first use of a real-time RT-PCR to quantify the amount of WNV in mosquito saliva.