腎症候性出血熱ウイルス抗体検査システムならびに汚染対応策の検討

1.克隆了编码汉滩病毒核衣壳蛋白的S基因组片段，并利用杆状病毒系统进行了表达。结论：1.成功地将表达的核衣壳蛋白作为抗原应用于ELISA试剂盒。确定试剂盒的基本条件及试剂盒的稳定性和效价，为试剂盒的研制奠定基础。从国立大学34个机构(1552份血清)、公立大学7个机构(454份血清)和私立大学22个机构(816份血清)中采集2822份实验大鼠血清样本，以检验该试剂盒的可靠性。在2822份血清标本中，有74份(2.6%)经肉眼判定后用酶联免疫吸附试验试剂盒检测为阳性。以汉滩和普玛拉汉坦病毒血清型为抗原，经IFA和Western印迹分析，所有阳性血清均为阴性，因此该试剂盒的特异性可达97%以上。讨论了《动物收容所预防肾综合征出血热指南(1981)》的修订情况。因此，提出如下建议：1)增加关于实验动物汉坦病毒感染的血清学诊断实用程序的新章节；2)增加关于汉坦病毒感染的血清学监测原理的说明。血清学诊断建议采用近期上市的ELISA试剂盒或委托其他公司进行临床诊断。新的或修订的指南将分发给相关设施。

1. The S genome segment which encodes nucleocapsid protein of Hantaan virus was cloned and expressed by baculovirus system. The expressed nucleocapsid protein was successfully applied as an antigen for ELISA kit.2. Basic conditions for the ELISA kit and stability and efficacy of the reagents were determined for establish ELISA kit for put it on the market.3. A total of 2,822 serum specimens from laboratory rats was obtained from 34 facilities of national university (1,552 sera), 7 facilities of public university (454 sera) and 22 facilities of private university (816 sera) to examine the reliability of the ELISA kit.4. Of the 2,822 serum specimens, 74 (2.6%) of them shown to be positive by the ELISA kit after naked eye decision. Since all of the positive sera were confirmed as negative by IFA and Western blot analysis with Hantaan and Puumala serotypes of hantaviruses as antigens, the specificity of the ELISA kit is considered as more than 97%.5. The revision of "Guideline for the prevention of hemorrhagic fever with renal syndrome amonglaboratory animal facility (Issued in 1981)" was discussed. As a results, following recommendation was made.1) Add the new chapter regarding the practical procedure for serologic diagnosis of hantavirus infection among laboratory animal.2) Add the description regarding the principle for serological monitoring for hantavirus infection. Serologic diagnosis is recommended to be done by ELISA kit that will be come on to the market in the near future or commission to other company for clinical diagnosis.6. The new or revised guideline will be distributed to related facilities.