Fujino Motoko, Yoshida Naoko, Kimura Keiko, Zhou Jianhui, Motegi Yoshie, Komase Katsuhiro, Nakayama Tetsuo
Kitasato Institutes for Life Sciences, Laboratory of Viral Infection, Shirokane 5-9-1, Minato-ku, Tokyo 108-8641, Japan.
J Virol Methods. 2007 Jun;142(1-2):15-20. doi: 10.1016/j.jviromet.2007.01.001. Epub 2007 Feb 23.
Sero-epidemiological studies are required to identify populations susceptible to measles. The hemagglutination inhibition (HI) test is no longer sensitive enough to confirm immunity to measles, and at present the particle agglutination (PA) test and enzyme-linked immunosorbent assay (EIA) are employed. The most reliable method is the neutralization test (NT), but it is time-consuming and requires experience. To simplify the NT, a recombinant measles AIK-C virus expressing green fluorescence protein (GFP-MVAIK) was constructed and used as a challenge virus. Plaques and cytopathic effects were visualized under ultraviolet light and detected easily, and measuring the intensity of the fluorescence enabled a reduction in the time-consuming steps. Neutralizing antibody titers of a complete inhibition neutralization test were equivalent to those of a 90% plaque reduction neutralization test. Comparison of four methods, HI, PA, EIA and the complete inhibition neutralization test, showed that only the results of EIA correlated well with those of the complete inhibition neutralization test, but sera with borderline levels by EIA were sometimes negative by the complete inhibition neutralization assay.
需要进行血清流行病学研究以确定对麻疹易感的人群。血凝抑制（HI）试验已不再足够灵敏以确认对麻疹的免疫力，目前采用颗粒凝集（PA）试验和酶联免疫吸附测定（EIA）。最可靠的方法是中和试验（NT），但该方法耗时且需要经验。为简化NT，构建了表达绿色荧光蛋白的重组麻疹AIK-C病毒（GFP-MVAIK）并用作攻击病毒。在紫外线下可观察到蚀斑和细胞病变效应且易于检测，并且通过测量荧光强度能够减少耗时步骤。完全抑制中和试验的中和抗体效价与90%蚀斑减少中和试验的效价相当。对HI、PA、EIA和完全抑制中和试验这四种方法的比较表明，只有EIA的结果与完全抑制中和试验的结果相关性良好，但EIA处于临界水平的血清有时在完全抑制中和试验中为阴性。
