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目的:介绍壳蛋白免疫法(hexonimmunoassay) ,并同传统细胞病变(CPE)法比较,检测其可靠性和可重复性。方法:3种重组腺病毒(Ad -lacz、Ad -hEndo和dl15 2 0 )扩增、提纯后,用壳蛋白免疫法、CPE法测定病毒滴度及A2 60 值法测定病毒总颗粒数。结果:壳蛋白免疫法和CPE法同时测定3种同批次病毒滴度,结果一致,P >0. 0 5 ;对不同批次3种腺病毒滴度测定,结果仍相似,平均差值 x±s=0. 15 2. 7±0 . 32 70 ,P >0 .0 5 ,两法有很好的相关性,r=0 913;壳蛋白免疫法对同批dl15 2 0测定19次,变异系数仅为1. 4 2 %。结论:壳蛋白免疫法是测定腺病毒滴度的有效方法,系统误差小、重复性好、结果客观可靠,可避免CPE法中由于细胞退化和工作人员主观判断造成的系统误差,比CPE法更精确、更省时。
OBJECTIVE: To introduce the hexonimmunoassay assay and compare it with the traditional cytopathic effect (CPE) method to test its reliability and repeatability. Methods: Three recombinant adenoviruses (Ad-lacz, Ad-hEndo and dl1520) were amplified and purified. The total virus counts were determined by the capsid protein immunoassay, the CPE method and the A260 value method. Results: The titers of three batches of viruses with the same batch assay were determined by the method of CPE and CPE simultaneously. The results were the same, P> 0.05. The titer of three kinds of adenovirus in different batches was still similar, the average difference x ± s = 0.15 2. 7 ± 0.32 70, P> 0 .0 5, there was a good correlation between the two methods, r = 0 913. The same assay was performed on the same batch of dl15 2 0 for 19 times, The coefficient of variation is only 1.42%. Conclusion: Enzyme immunoassay is an effective method for the determination of adenovirus titer. The system error is small and the repeatability is good. The results are objective and reliable, which can avoid systematic errors caused by cell degeneration and staff subjective judgment in CPE method. Accurate, more time-saving.