Warnes A, Fooks A R, Stephenson J R
Biologics Division, PHLS, CAMR, Porton Down, Salisbury, UK.
J Virol Methods. 1994 Oct;49(3):257-68. doi: 10.1016/0166-0934(94)90141-4.
Measles nucleoprotein has been successfully expressed in three different hosts, bacterial (Escherichia coli BL21 DE3), insect (Spodoptera frugiperda; Sf9) and mammalian (primary human fibroblasts) cells, each producing an antigenic protein of M(r) 60 kDa. The nucleoprotein produced in all three hosts was used in an ELISA for the detection of antibodies to measles virus in a cohort of haemagglutinin-positive or -negative human sera. Data produced from baculovirus and adenovirus-based antigens indicated that there was good correlation between the ELISA results and previous haemagglutination inhibition test data, and there was little background interference by cellular proteins or the development of false positive or negative results. The assay was rapid as it could be carried out in under 4 h, sensitive as the sera could be diluted by at least 1000-fold, and versatile as both IgG and IgM could be detected and differentiated.
麻疹核蛋白已在三种不同宿主中成功表达，分别是细菌（大肠杆菌BL21 DE3）、昆虫（草地贪夜蛾；Sf9）和哺乳动物（原代人成纤维细胞）细胞，每种宿主都产生了分子量为60 kDa的抗原性蛋白。在酶联免疫吸附测定（ELISA）中，使用在所有这三种宿主中产生的核蛋白来检测一组血凝素阳性或阴性人血清中的麻疹病毒抗体。基于杆状病毒和腺病毒的抗原所产生的数据表明，ELISA结果与先前的血凝抑制试验数据之间具有良好的相关性，并且细胞蛋白几乎没有背景干扰，也未出现假阳性或假阴性结果。该检测方法快速，因为它可在4小时内完成；灵敏，因为血清可至少稀释1000倍；通用，因为可检测和区分IgG和IgM。
