含基孔肯雅病毒包膜蛋白VSV假病毒制备

Preparation of pseudotype VSV with chikungunya virus envelop protein

目的建立一种含有基孔肯雅病毒(CHIKV)包膜蛋白的便于流行病学调查的疱疹性口炎病毒(VSV)假病毒。方法以缺失VSV G蛋白的VSV病毒做为载体,制备含有CHIKV膜蛋白的VSV-E2E1病毒。结果该VSV假病毒带有CHIKV膜蛋白,同时带有绿色荧光蛋白(GFP)或荧光素酶;功效性试验显示,VSV-E2E1病毒在CHIKV E2单克隆抗体1:2 000稀释度时,酶的活性为2.73%,1:4 000稀释度时酶活性为9.61%,1:8 000稀释度时酶活性为19.76%,1:16 000稀释度时酶活性为29.23%,1:32 000稀释度时酶活性为41.68%,1:64 000稀释度时酶活性为74.84%;无病毒感染的阴性对照酶活性为2.46%,当抗体稀释度为1:2 000时,最大限度地抑制了病毒的复制。结论含有基孔肯雅病毒包膜蛋白VSV假病毒安全、可靠,可应用于流行病学调查分析。

Objective To develope a method for the preparation of pseudotype vesicular stomatitis virus （VSV） with chikungunya virus（ CHIKV） envelop protein. Methods VSV without G protein genome was used as the vector to produce pseudotype VSV. Results Pseudotype VSV with CHIKV envelop protein and green fluorescent protein （GFP） was developed. In the efficacy test,the infectivity of the pseudotype VSV constructed was 2. 73% for the 1：2 000 dilution of the antibody against CHIKV E2 of VSV-E2E1 virus, 9. 61% for the dilution of 1.4 000,19. 76% for the dilution of 1：8 000,29. 23% for the dilution of 1：16 000,41.68% for the dilution of 1：32 000,and 74. 84% for the dilution of 1 ： 64 000, respectively; the infectivity of negative control was 2. 46 %. The viral replication was mostly inhibited under the dilution of 1：2 000 for the antibody against CHIKV E2. Conclusion The constructed pseudotype VSV is safe and stable and can be applied in epidemic survey by measuring the expression of GFP in infected cells or the activity of luciferase.