HIV假病毒用于p24抗原检测试剂研制的研究

Research on HIV Pseudovirion as Positive Control for p24 Antigen Detecting Kit

为了解决在研制新型HIV检测试剂盒时遇到的p24蛋白阳性对照物保存不便、阳性对照物稳定性不高以及蛋白结构和聚集形式与天然HIV病毒的p24存在差异等问题,通过将改造的HIV-1骨架质粒pNL43 EGFP R-E-和包膜蛋白质粒pGX-C共转染293T细胞后分离细胞培养液,获得具有单轮感染活性的HIV-1假病毒。通过对此假病毒颗粒的天然衣壳蛋白p24和市售HIV检测试剂盒中的p24蛋白阳性对照物的阳性反应有效性和保存稳定性的检测,证明了HIV-1假病毒p24蛋白具有更易保存、更稳定、更接近天然病毒p24蛋白的优点,适宜作为HIV检测试剂盒阳性对照物。

A new kind of p24 antigen positive control was developed to provide a better solution to the problems encountered in developing a novel HIV detecting kit, such as （a） inconveniency in storing p24 positive control, （b） difficulty in maintaining p24 antigen activity, （c） differences in protein structure and aggregation mode between recombinant p24 antigen and natural p24 antigen. Modified HIV-1 skeleton plasmid pNL43 EGFP R-E- and envelope protein plasmid pGX-C were co-transfccted into 293T cell, and then culture fluid containing single-round HIV pseudovirions were collected. This kind of HIV pseudovirions possess natural core shell protein p24, which could be used as the positive control for detection of HIV p24 antigen. Through comparison of activity and stability between pseudovirion p24 antigen and positive control in common detecting kit, it was proved that pseudovirion p24 is more convenient to store, more stable and more similar to natural p24 protein structure and aggregation mode than common recombinant p24 protein.