Obara Mayumi, Yoshii Kentaro, Kawata Tomoko, Hayasaka Daisuke, Goto Akiko, Mizutani Tetsuya, Kariwa Hiroaki, Takashima Ikuo
Laboratory of Public Health, Department of Environmental Veterinary Sciences, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo 060-0818, Japan.
J Virol Methods. 2006 Jun;134(1-2):55-60. doi: 10.1016/j.jviromet.2005.11.018. Epub 2006 Mar 15.
The similarity of symptoms produced by tick-borne encephalitis (TBE) and Japanese encephalitis (JE) and the high degree of cross-reactivity between TBE and JE viruses by serological tests make the development of a differential diagnostic test a priority. In this study, recombinant prM/E proteins of TBE virus strain Oshima 5-10 expressed in mammalian cells resulted in the release of subviral particles (SPs) into the culture medium. Using the SPs as antigens, enzyme-linked immunosorbent assay (ELISA) systems were developed to detect TBE virus-specific IgM and IgG antibodies, designated SP-IgG and SP-IgM ELISAs, respectively. Of 83 serum samples from encephalitis patients in Khabarovsk, Russia, which were positive with the neutralization test (NT), 82 were positive by the SP-IgG ELISA, for a sensitivity of 98.8%, which was higher than that of a commercial ELISA kit. All 12 NT-negative samples were also negative by the SP-IgG ELISA (specificity, 100%). Of 17 patient samples that were NT-positive, 16 (94.1%) were positive by the SP-IgM ELISA. Of 15 paired serum samples that yielded equivocal results by NT, 11 had positive results with the SP-IgM ELISA, indicating a diagnosis of TBE infection. The SP-IgG and SP-IgM ELISAs showed no cross-reactivity with antibodies to the JE virus. The results indicate that these ELISAs will be useful for the detection of TBE-specific antibodies.
蜱传脑炎（TBE）和日本脑炎（JE）产生的症状相似，且通过血清学检测发现TBE病毒和JE病毒之间存在高度交叉反应，因此开发一种鉴别诊断测试成为当务之急。在本研究中，在哺乳动物细胞中表达的TBE病毒大岛5 - 10株的重组prM/E蛋白导致亚病毒颗粒（SPs）释放到培养基中。以SPs作为抗原，开发了酶联免疫吸附测定（ELISA）系统来检测TBE病毒特异性IgM和IgG抗体，分别命名为SP - IgG ELISA和SP - IgM ELISA。在俄罗斯哈巴罗夫斯克的83份脑炎患者血清样本中，中和试验（NT）呈阳性，其中82份通过SP - IgG ELISA检测呈阳性，灵敏度为98.8%，高于商用ELISA试剂盒。所有12份NT阴性样本通过SP - IgG ELISA检测也为阴性（特异性为100%）。在17份NT阳性的患者样本中，16份（94.1%）通过SP - IgM ELISA检测呈阳性。在15份通过NT产生不确定结果的配对血清样本中，11份通过SP - IgM ELISA检测呈阳性，表明诊断为TBE感染。SP - IgG和SP - IgM ELISA与JE病毒抗体无交叉反应。结果表明，这些ELISA将有助于检测TBE特异性抗体。
