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. 利用杆状病毒系统克隆并表达了汉滩病毒编码核衣壳蛋白的S基因组片段。成功地将表达的核衣壳蛋白作为抗原应用于ELISA试剂盒。2 .确定酶联免疫吸附测定试剂盒的基本条件及各试剂的稳定性和有效性，建立酶联免疫吸附测定试剂盒，并将其推向市场。从34所国立大学（1552份）、7所公立大学（454份）和22所私立大学（816份）共采集实验大鼠血清2822份，检验ELISA试剂盒的可靠性。在2822份血清标本中，74份（2.6%）经肉眼检测后ELISA试剂盒呈阳性。由于所有阳性血清均以汉坦病毒的汉坦和普马拉血清型为抗原，经IFA和Western blot分析均为阴性，因此认为ELISA试剂盒的特异性大于97% 5。对《实验室动物预防肾综合征出血热指南（1981年）》的修订进行了讨论。因此，提出了以下建议。1)新增实验动物汉坦病毒感染血清学诊断实用程序一章。2)增加汉坦病毒感染血清学监测原理说明。建议采用ELISA试剂盒进行血清学诊断，该试剂盒将于近期上市或委托其他公司进行临床诊断。新的或经修订的指引将分发给有关设施。

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.