Dearden C J, Al-Nakib W
MRC Common Cold Unit, Harvard Hospital, Salisbury, Wiltshire, England.
J Med Virol. 1987 Oct;23(2):179-89. doi: 10.1002/jmv.1890230211.
This paper describes the first enzyme-linked immunosorbent assay for the detection of rhinovirus antigens in clinical specimens (nasal washings), either directly or following overnight cell culture amplification. The assay takes approximately 48 hours to perform and utilizes the same rabbit antirhinovirus hyperimmune serum as both the capture and detecting antibody. The latter has been biotin-labelled and is detected via a streptavidin beta-galactosidase preformed complex. This new assay has been found to be very sensitive, detecting human rhinovirus (HRV)-EL and HRV-2 at titres as low as 10(1.8) TCID50 100 microliter-1 and less than 10(1)TCID50 100 microliter -1, respectively. Furthermore, when 57 different human rhinovirus serotypes were tested in both the HRV-EL and HRV-2 ELISA systems a total of 49 (86%) were found to be cross-reactive. Of 36 clinical specimens tested by virus isolation, cell-culture-amplified (CCA) ELISA, and direct ELISA, 15 were positive by isolation, 11 by CCA-ELISA, and 11 by direct ELISA. The overall correlation of the CCA and direct ELISA techniques with virus isolation was found to be 88.9% and 66.7%, respectively. The present study demonstrates that the ELISA system developed is a sensitive technique for the diagnosis of rhinovirus infections.
本文描述了第一种用于检测临床标本（洗鼻液）中鼻病毒抗原的酶联免疫吸附测定法，该方法可直接检测，也可在过夜细胞培养扩增后进行检测。该测定法大约需要48小时完成，并且使用同一种兔抗鼻病毒超免疫血清作为捕获抗体和检测抗体。后者已用生物素标记，并通过链霉亲和素β-半乳糖苷酶预形成复合物进行检测。已发现这种新测定法非常灵敏，分别能检测到低至10(1.8)TCID50 100微升-1的人鼻病毒（HRV）-EL和低于10(1)TCID50 100微升-1的HRV-2。此外，当在HRV-EL和HRV-2酶联免疫吸附测定系统中对57种不同的人鼻病毒血清型进行检测时，共发现49种（86%）有交叉反应。在通过病毒分离、细胞培养扩增（CCA）酶联免疫吸附测定法和直接酶联免疫吸附测定法检测的36份临床标本中，病毒分离阳性的有15份，CCA酶联免疫吸附测定法阳性的有11份，直接酶联免疫吸附测定法阳性的有11份。发现CCA和直接酶联免疫吸附测定技术与病毒分离的总体相关性分别为88.9%和66.7%。本研究表明，所开发的酶联免疫吸附测定系统是诊断鼻病毒感染的一种灵敏技术。
