Development of new serodiagnostic method for hemorrhagic fever with renal syndrome(HFRS) and vaccine development against HFRS.

在过去的三年里，我们致力于肾综合征出血热(HFRS)病毒的血清学诊断方法的研究和HFRS灭活疫苗的初步研究。1)新的HFRS诊断方法的开发；免疫荧光(IF)技术已用于检测人和啮齿动物血清中的HFRS病毒抗体。我们开始开发新的血清学诊断方法，如中和抗体(NT)试验、血凝抑制(HI)试验和ELISA试验。首先，用过氧化物酶-抗过氧化物酶染色计数肾综合征出血热病毒引起的病灶数。这种方法比斑块形成法更便于NT试验。其次，从肾综合征出血热病毒感染的细胞液中制备血凝抗原，用于血凝抑制试验。血凝抑制试验显示各毒株间有较强的交叉反应。最后建立了抗体捕获ELISA法，并比较了IF、HI和ELISA法检测抗体的效果。酶联免疫吸附试验和免疫荧光试验比血凝抑制试验更敏感，但血凝抑制试验比其他两种试验更容易。2)疫苗的研制：我们已经分离出用于分析肾综合征出血热病毒抗原的单抗。病毒粒子上有两个糖蛋白(G1、G2)，它们与NT和HI活性相关。接下来，将不同的HFRS病毒接种到新生的小鼠或大鼠身上，可以发现HFRS病毒的毒力是不同的，这取决于受感染动物的种类以及病毒。最后，从小鼠脑内制备灭活疫苗，并用该灭活疫苗免疫小鼠，诱导产生NT抗体。在进一步的动物实验后，可望作为人和啮齿动物的疫苗。

We have been engaged for the last three years in the development of the method for the serodiagnosis of hemorrhagic fever with renal syndrome(HFRS) virus, and also on the preliminary study of inactivated vaccine against HFRS.1)development of new diagnostic method for HFRS; The immunofluorescent (IF) technique has been used to detect antibody against HFRS virus in human and rodent sera. We started to develop new serodiagnostic methods such as neutralizing antibody (NT) test, hemagglutination inhibition (HI) test, and ELISA test. First, peroxidase-antiperoxidase staining was applied for counting foci induced by HFRS virus. This method is more convenient than plaque formation method for NT test. Second, hemagglutinating antigen was prepared from HFRS virus-infected cell fluid, and this antigen was applied for HI test. Strong cross-reactions among various virus strains were demonstrated by HI test. Finally, antibody captured ELISA test was developed, and IF, HI and ELISA tests were compared to detect antibodies. ELISA and IF tests were more sensitive than HI test, but HI test was easier than other two.2)development of vaccine: we have isolated monoclonal antibodies to analyse antigens of HFRS viruses. Two glycoproteins(G1, G2) were indentified on the viral virion, which correlated with NT and HI activities. Next, various HFRS viruses were inoculated to newborn mice or rats, and it could be found that the virulence of HFRS viruses is variable, depending on the species of infected animals as well as the viruses. Finally, inactivated vaccine was prepared from mice brains, and NT antibodies were induced in mice immunized with vaccine. It can be expected to be used as a vaccine for human and rodent after further experiments in animals.