Virus Research Center, Clinical Research Division, Sendai Medical Center, Sendai 983-8520, Japan.
J Virol Methods. 2010 Mar;164(1-2):24-9. doi: 10.1016/j.jviromet.2009.11.019. Epub 2009 Nov 17.
To apply serological testing for human metapneumovirus (hMPV) to large-scale sera samples, an enzyme-linked immunosorbent assay (ELISA) was developed in which purified virions were used as the antigen. The ELISA was evaluated using 102 human sera specimens from patients aged 0-59 years. There was a positive association between the ELISA results and neutralization test titers, with the correlation coefficients being greater in children <6 years old (rho=0.899, P<0.0001), which is consistent with a primary infection, than in persons >or=6 years old (rho=0.523, P<0.0001). Fifty sera samples were subjected to radioimmunoprecipitation to measure the quantity of antibodies to the fusion protein (RIP-F) and the nucleoprotein (RIP-N). The results showed significant associations between the ELISA titers and the amount of RIP-F as determined by radioimmunoprecipitation in children <6 years old (rho=0.804, P=0.0083) and in persons >or=6 years old (rho=0.577, P=0.0009). The correlation between the ELISA titer and the amount of RIP-N determined by radioimmunoprecipitation was not significant in persons >or=6 years old (rho=0.417, P=0.0829), although this correlation was significant in children <6 years old (rho=0.764, P=0.0137). The ELISA titer correlated with the amount of antibodies to the F protein, but not to the N protein. This whole virus-based ELISA will be useful for the diagnosis of hMPV infection in clinical laboratories and is also useful for the large-scale investigations, such as seroprevalence among residents of a particular region.
为了将血清学检测人类偏肺病毒（hMPV）应用于大规模的血清样本，我们开发了一种酶联免疫吸附试验（ELISA），其中使用纯化的病毒粒子作为抗原。使用 102 份年龄在 0 至 59 岁的患者血清标本对 ELISA 进行了评估。ELISA 结果与中和试验滴度之间存在正相关，6 岁以下儿童的相关系数更高（rho=0.899，P<0.0001），这与初次感染一致，而 6 岁以上人群的相关系数较低（rho=0.523，P<0.0001）。50 份血清样本用于放射免疫沉淀法测量融合蛋白（RIP-F）和核蛋白（RIP-N）的抗体量。结果表明，在 6 岁以下儿童（rho=0.804，P=0.0083）和 6 岁以上人群（rho=0.577，P=0.0009）中，ELISA 滴度与 RIP-F 的量之间存在显著相关性。在 6 岁以上人群中，ELISA 滴度与 RIP-N 的量之间的相关性不显著（rho=0.417，P=0.0829），尽管在 6 岁以下儿童中，这种相关性显著（rho=0.764，P=0.0137）。ELISA 滴度与 F 蛋白的抗体量相关，但与 N 蛋白不相关。这种基于整个病毒的 ELISA 将有助于临床实验室对 hMPV 感染的诊断，也有助于大规模调查，如特定地区居民的血清流行率。
