Institute for Animal Experimentation, Faculty of Medicine, Hokkaido University, Kita 15, Nishi 7, Kita-ku, Sapporo, Hokkaido 060-8638, Japan.
ICLAS Monitoring Center, Central Institute for Experimental Animals, 3-25-12 Tonomachi, Kawasaki, Kanagawa 210-0821, Japan.
Exp Anim. 2021 May 13;70(2):161-168. doi: 10.1538/expanim.20-0099. Epub 2020 Nov 12.
Rapid and simple serologic tests that require only a small amount of blood without the euthanization of animals are valuable for microbial control in colonies of laboratory animals. In this study, we developed a multiplex immunochromatographic assay (ICA) for detection of antibodies to Sendai virus (also known as hemagglutinating virus of Japan), hantavirus, and sialodacryoadenitis virus, which are causative agents of major infectious diseases in rats. For this assay, an ICA strip was placed into a microtube containing 150 µl PBS and either 0.75 µl of rat serum or 1.5 µl of whole blood. Binding antibodies were visualized by using anti-rat IgG antibody-conjugated colloidal gold. Under these conditions, the multiplex ICA simultaneously and specifically detected antibodies to multiple antigens. Positive serum samples for each infectious disease were used to evaluate the sensitivity and specificity of the multiplex ICA. The sensitivities of the multiplex ICA for Sendai virus, hantavirus, and sialodacryoadenitis virus were 100%, 100%, and 81%, respectively. No nonspecific reactions were observed in any of the 52 positive sera against heterologous antigens. In addition, 10 samples of uninfected sera did not show any bands except for the control line. These observations indicate high specificity of the multiplex ICA. Moreover, the multiplex ICA could be applied to diluted blood. These results indicate that the multiplex ICA is appropriate for rapid and simple serological testing of laboratory rats.
快速且简单的血清学检测方法，只需少量血液，无需处死动物，对于控制实验动物群体中的微生物非常有价值。在这项研究中，我们开发了一种用于检测仙台病毒（也称为日本血凝病毒）、汉坦病毒和涎泪腺炎病毒抗体的多重免疫层析检测法（ICA），这些病毒是导致大鼠主要传染病的病原体。对于该检测方法，将 ICA 条放入含有 150 µl PBS 的微管中，管内分别含有 0.75 µl 大鼠血清或 1.5 µl 全血。通过使用抗大鼠 IgG 抗体偶联胶体金来可视化结合抗体。在这些条件下，多重 ICA 可同时特异性地检测多种抗原的抗体。使用每种传染病的阳性血清样本评估了多重 ICA 的灵敏度和特异性。该多重 ICA 对仙台病毒、汉坦病毒和涎泪腺炎病毒的灵敏度分别为 100%、100%和 81%。在针对异源抗原的 52 份阳性血清中，未观察到任何非特异性反应。此外，除控制线外，10 份未感染血清样本均未显示任何条带。这些观察结果表明多重 ICA 具有很高的特异性。此外，该多重 ICA 可应用于稀释的血液。这些结果表明，该多重 ICA 适用于快速且简单的实验大鼠血清学检测。
