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比较双抗体夹心ELISA法与直接免疫荧光(DFA)法在检测汉坦病毒灵敏度方面的差别。用汉坦病毒76—118株感染Vero—E6细胞,取不同时间点的病毒感染细胞制备细胞爬片或细胞培养物冻融上清,分别用本室制备的抗汉坦病毒单克隆抗体标记荧光素或辣根过氧化物酶(HRP),建立直接免疫荧光法和双抗体夹心ELISA法检测上述细胞中的病毒抗原,并比较两者的检测灵敏度。用空斑形成实验结果作为金标准,以评价两种方法在检测病毒抗原灵敏度方面的差异。结果:用两种检测方法测定不同时间点的汉坦病毒培养物抗原,双抗体夹心ELISA法不仅检出的时间早,而且其病毒滴度均较IFA法高(P<0.01)。说明ELISA法检测汉坦病毒滴度的灵敏度较IFA法更高,并且操作简单、可重复性好、便于大批量规模化检测,因此更适用于汉坦病毒的检测。
The difference between the double antibody sandwich ELISA method and the direct immunofluorescence (DFA) method in the detection of Hantavirus sensitivity was compared. Vero-E6 cells were infected with Hantavirus strain 76-118 and virus-infected cells at different time points were used to prepare cell slide or freeze-thaw supernatant of cell culture. The anti-Hantaan virus monoclonal antibody labeled in this chamber was used to label the fluorescence (HRP), direct immunofluorescence and double antibody sandwich ELISA were used to detect the virus antigens in the above cells, and the detection sensitivity was compared between the two. Plaque formation experiments were used as the gold standard to assess differences in the sensitivity of the two methods in detecting viral antigens. Results: The antigens of Hantavirus culture at different time points were detected by two methods. The double antibody sandwich ELISA not only detected the early time but also had higher virus titers than IFA method (P <0.01). This indicated that the sensitivity of ELISA method for detecting Hantavirus titer was higher than that of IFA method, and its operation was simple and the repeatability was good. It was more suitable for mass scale detection and was more suitable for the detection of Hantavirus.