Duvdevani P, Varsano N, Slepon R, Lerman Y, Shohat T, Mendelson E
Central Virology Laboratory, Chaim Sheba Medical Center, Tel-Hashomer, Israel.
Clin Diagn Virol. 1996 Oct;7(1):1-6. doi: 10.1016/s0928-0197(96)00240-1.
Determination of the immune status against measles in young adults requires careful evaluation of the laboratory methods because of waning immunity. The hemagglutination inhibition (HI) test and enzyme-linked immunosorbent assay (ELISA) may lack the sensitivity required to detect very low levels of antibodies. In addition, the correlation between ELISA-IgG assays and the degree of protection from measles is not well defined.
(a) Evaluation of a commonly used measles ELISA-IgG test kit in comparison with the hemagglutination inhibition (HI) test which corresponds strongly to virus neutralization; (b) determination of false negative rates of the ELISA-IgG and the HI tests; (c) evaluation of the ELISA-IgG test kit as a quantitative assay.
One hundred and eighty serum samples collected from 60 vaccinated young adults immediately before vaccination and 14 and 28 days postvaccination, were tested comparatively by HI and by a commercial ELISA-IgG kit. For evaluation of false negative rates, postvaccination sera of a cohort of 48 vaccinees with negative HI or ELISA-IgG prevaccination sera were tested for IgM. Sixty-three of the samples were also titrated by the ELISA-IgG kit using serial dilutions, for comparison with HI titers.
Using the HI test as a reference method, the ELISA-IgG kit was found to have overall accuracy of 81%, sensitivity of 80% and specificity of 84%. The false negative and the false positive rates were 20% and 16%, respectively. In contrast, when we used postvaccination IgM test to distinguish between true and false prevaccination negatives in both the HI and ELISA-IgG tests, we found that the false negative rates were 75.6% by ELISA and 72.5% by HI, and false positive rates were 2.4% and 0%, respectively. Serum titers determined by the ELISA-IgG test were generally 5-10-fold higher than the corresponding HI titers, but without a consistent correlation.
Both the ELISA-IgG and the HI tests frequently failed to detect residual immunity. The two tests also did not correlate well with each other suggesting that different antigenic determinants of the virus are involved in each assay and therefore the HI test should not be used as a reference method for evaluation of the sensitivity of ELISA IgG kits.
由于免疫力下降,在年轻成年人中测定麻疹免疫状态需要仔细评估实验室方法。血凝抑制(HI)试验和酶联免疫吸附测定(ELISA)可能缺乏检测极低水平抗体所需的灵敏度。此外,ELISA-IgG测定与麻疹保护程度之间的相关性尚不明确。
(a)将一种常用的麻疹ELISA-IgG检测试剂盒与血凝抑制(HI)试验进行比较评估,HI试验与病毒中和密切相关;(b)确定ELISA-IgG和HI试验的假阴性率;(c)评估ELISA-IgG检测试剂盒作为定量测定方法。
从60名接种疫苗的年轻成年人中收集180份血清样本,分别在接种疫苗前、接种后14天和28天进行采集,通过HI试验和一种商用ELISA-IgG试剂盒进行比较检测。为了评估假阴性率,对48名接种者的接种后血清进行检测,这些接种者接种前的HI或ELISA-IgG血清为阴性,检测其IgM。其中63份样本还用ELISA-IgG试剂盒进行系列稀释滴定,以与HI滴度进行比较。
以HI试验作为参考方法,发现ELISA-IgG试剂盒的总体准确率为81%,灵敏度为80%,特异性为84%。假阴性率和假阳性率分别为20%和16%。相比之下,当我们使用接种后IgM试验来区分HI和ELISA-IgG试验中接种前真阴性和假阴性时,我们发现ELISA的假阴性率为75.6%,HI为72.5%,假阳性率分别为2.4%和0%。ELISA-IgG试验测定的血清滴度通常比相应的HI滴度高5-10倍,但没有一致的相关性。
ELISA-IgG和HI试验都经常无法检测到残余免疫力。这两种试验相互之间的相关性也不好,这表明病毒的不同抗原决定簇参与了每种试验,因此HI试验不应作为评估ELISA IgG试剂盒灵敏度的参考方法。