柯萨奇病毒A组16型中和抗体假病毒荧光定量检测方法的建立及初步应用

发布时间：2018-04-27 07:19

本文选题：柯萨奇病毒A型 + 假病毒　；参考：《中国病毒病杂志》2016年01期

【摘要】：目的建立一种简单、快速、高通量的检测柯萨奇病毒A组16型(CV-A16)中和抗体假病毒荧光定量检测方法 (pseudovirus luciferase assay,PVLA)。方法首先构建CV-A16核衣壳蛋白表达子及插入萤火虫荧光素酶报告基因的肠道病毒71型(EV-A71)复制子,再顺次转染293T细胞,互补包装得到CVA16假病毒。通过条件优化并使用CV-A16国家中和抗体标准品,建立CV-A16中和抗体定量检测方法(PVLA)。用该方法检测15份CV-A16病毒免疫小鼠血清,并与传统CPE法的检测结果进行统计学分析,比较两种方法检测结果的一致性。结果得到CV-A16假病毒,建立了CV-A16假病毒中和抗体检测方法,该方法检测CV-A16中和抗体滴度的结果与传统CPE法的检测结果高度一致(相关系数r2为0.91)。结论CV-A16假病毒中和抗体检测方法可作为CPE法的替代方法,用于CV-A16中和抗体的快速检测。
[Abstract]:Objective to establish a simple, rapid and high throughput method for the detection of pseudovirus luciferase assayayae of coxsackie virus A group 16 (CV-A16). Methods CV-A16 core-capsid protein expression and Enterovirus 71 EV-A71) replicon inserted into luciferase reporter gene of firefly were constructed, and then transfected into 293T cells in sequence. CVA16 pseudoviruses were obtained by complementary packaging. By optimizing the conditions and using the neutralizing antibody standard of CV-A16, a quantitative detection method for neutralizing antibody of CV-A16 was established. Fifteen sera of mice immunized with CV-A16 virus were detected by this method and compared with the results of traditional CPE method. Results CV-A16 pseudovirus was obtained and a method for detecting CV-A16 neutralizing antibody was established. The results of CV-A16 neutralization antibody detection by this method were in good agreement with those of the traditional CPE method (correlation coefficient r 2 was 0.91g / kg). Conclusion the method of CV-A16 pseudovirus neutralizing antibody can be used as an alternative to CPE method for rapid detection of CV-A16 neutralizing antibody.
【作者单位】：中国食品药品检定研究院肝炎病毒疫苗室;华兰生物工程股份有限公司研发部;
【基金】：国家科技重大专项(2012ZX10004701)
【分类号】：R392

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