西尼罗病毒空斑形成试验方法的建立与应用

目的建立西尼罗病毒的空斑形成试验方法，应用于病毒滴度测定和实验感染蚊虫及来亨鸡血液样本中病毒的定量检测。方法将稀释的样本接种常规制备的Veto细胞单层，用琼脂糖凝胶覆盖，孵育一定时间后，加中性红染色，计数空斑数，计算空斑形成单位。结果在琼脂糖凝胶中西尼罗病毒可形成直径大约1～3mm的圆形或类圆形空斑。20％病毒鼠脑悬液的病毒滴度为10^7空斑形成单位。感染蚊虫样本和来亨鸡血液样本也出现典型的空斑。结论建立了西尼罗病毒的空斑形成试验方法，该方法可应用于病毒滴度测定和实验感染蚊虫及来亨鸡血液样本中病毒的定量检测。

Establishment and Application of Plaque Assay for the Detection of West Nile Virus

Objective To establish plaque assay for the quantitative detection of West Nile virus(WNV). Methods Vero cell was selected to be infected with WNV. Approximately 24 h before beginning the titration protocol, Vero cells were plated in 6-well plates to form an even monolayer. Virus in specimens was serially diluted and added to each well to infect the cells. The plates were covered with agarose overlay medium and incubated in an incubator. The plates were stained by a solution of Neutral red, and the number of plaques was counted. Results The plaques appeared as clear circles( 1-3 mm) against a red or pink background. The quantity of virus in infected mouse brain tissues was 10~7 pfu. Conclusion The plaque assay is confirmed as an efficient and rapid protocol of quantitatively detecting WNV in experimentally infected mosquitoes and Leghorn chicken.