A reverse-type sandwich enzyme-linked immunosorbent assay for detecting antibodies to Borna disease virus
A reverse-type sandwich enzyme-linked immunosorbent assay for detecting antibodies to Borna disease virus
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反向夹心酶联免疫吸附法检测博尔纳病病毒抗体
DOI：
10.1128/jcm.35.7.1661-1666.1997
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发表时间：
1997
影响因子：
9.4
通讯作者：
Masato Tashiro
中科院分区：
文献类型：
--
作者：
Taisuke Horimoto;Hiromi Takahashi;Masahiro Sakaguchi;Kaori Horikoshi;Shuji Iritani;Hajime Kazamatsuri;Kazuhiko Ikeda;Masato Tashiro
为探讨博尔纳病病毒（Borna disease virus，BDV）感染与神经精神疾病的流行病学关系，建立了检测BDV特异性抗体的反向夹心酶联免疫吸附试验（RS-ELISA）。在该试验中，用BDV p40抗原分散包被微孔板威尔斯孔，然后加入低稀释度的供试品，然后加入生物素化的p40。使用预先形成的链霉亲和素和辣根过氧化物酶结合的生物素和酶底物的复合物来测量捕获的生物素化的p40。理论上，RS-ELISA应该特异性地检测抗BDV抗体而没有非特异性信号;这样的信号可能在常规血清学测定中出现。此外，RS-ELISA可以在相同的方案下应用于测试来自各种动物的样品。用抗BDV大鼠和兔血清对该方法进行了标准化，使其具有较高的特异性和敏感性。当我们使用RS-ELISA测定日本东京地区70名慢性精神分裂症患者和40名健康个体血浆中抗BDV抗体的存在时，没有发现血浆样品具有针对BDV p40的特异性抗体，表明在我们的测试人群中BDV感染与疾病之间没有关联。先前通过免疫荧光或免疫印迹试验判定为BDV血清阳性的血清也显示阴性反应。这些结果表明，由于非特异性反应引起的感染假阳性病例可能包括在先前关于BDV的血清流行病学信息中。
To investigate whether there is an epidemiological correlation between Borna disease virus (BDV) infection and human neuropsychiatric diseases, we established a reverse-type sandwich enzyme-linked immunosorbent assay (RS-ELISA) for detecting specific antibodies to BDV. In this assay, microplate wells were coated dispersely with BDV p40 antigen, followed by the addition of test samples at a low dilution and then the biotinylated p40. A preformed complex of streptavidin and horseradish peroxidase-conjugated biotin and an enzyme substrate were used to measure the captured biotinylated p40. Theoretically, RS-ELISA should specifically detect anti-BDV antibodies without nonspecific signals; such signals possibly occur in conventional serological assays. Additionally, the RS-ELISA could be applied under the same protocols to test samples from a variety of animals. By using anti-BDV rat and rabbit sera, the assay was standardized so that it had high specificity and sensitivity. When we used the RS-ELISA to determine the presence of anti-BDV antibodies in plasma from 70 patients with chronic schizophrenia as well as 40 healthy individuals in the Tokyo area of Japan, no plasma sample was found to possess specific antibodies to BDV p40, indicating no association between BDV infection and the disease in our testing population. A negative reaction was also shown for the sera that had previously been judged to be seropositive for BDV by an immunofluorescence or immunoblot test. These findings suggested that false-positive cases of infection due to nonspecific reactions may be included in previous seroepidemiological information with regard to BDV.