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首页> 外文期刊>Clinical and diagnostic laboratory immunology >Simultaneous Serodetection of 10 Highly Prevalent Mouse Infectious Pathogens in a Single Reaction by Multiplex Analysis
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Simultaneous Serodetection of 10 Highly Prevalent Mouse Infectious Pathogens in a Single Reaction by Multiplex Analysis

机译：多重分析同时检测单个反应中10种高度流行的小鼠感染性病原体

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Under current practices of mouse colony maintenance, sera from mice are analyzed for antibodies against several widespread infectious pathogens by conventional immunoassays, generally enzyme-linked immunosorbent assay (ELISA). To test for multiple agents, these methods consume large volumes of mouse serum and are laborious and time-consuming. More efficient immunoassays, using small amounts of sample, are therefore needed. Accordingly, we have developed a novel multiplex diagnostic system that employs fluorescent microbeads, coated with purified antigens, for simultaneous serodetection of 10 mouse infectious agents. Individually identifiable, fluorescent microbeads were coated with antigens from Sendai virus, mouse hepatitis virus, Theiler's mouse encephalomyelitis virus/GDVII strain, mouse minute virus, mouse cytomegalovirus, respiratory enteric orphan virus (Reo-3 virus), mouse parvovirus, calf rotavirus for epizootic diarrhea virus of infant mice, vaccinia virus for ectromelia virus, and Mycoplasma pulmonis. Standard sera, singly positive for antibodies to individual infectious agents, were generated by inoculation of BALB/cj and C57BL/6j mice. Sera from these experimentally infected mice, as well as sera from naturally infected mice, were analyzed using a mixture of microbeads coated with antigens of the 10 infectious agents listed above. Results demonstrated that the multiplex assay was at least as sensitive and specific as ELISA for serodetection. Importantly, the multiplex assay required only 1 microliter of serum for simultaneous serodetection of the 10 mouse infectious agents in one reaction vessel. Thus, this multiplex microbead assay is a reliable, efficient, and cost-effective diagnostic modality that will impact serosurveillance of mice used in research.
机译：在小鼠集落维持的当前实践下，通过常规免疫测定法，通常是酶联免疫吸附测定法（ELISA），分析小鼠血清中针对几种广泛的传染性病原体的抗体。为了测试多种试剂，这些方法消耗了大量的小鼠血清，既费力又费时。因此，需要使用少量样品的更有效的免疫测定。因此，我们已经开发出一种新颖的多重诊断系统，该系统采用涂有纯化抗原的荧光微珠同时对10种小鼠传染原进行血清学检测。可分别鉴定的荧光微珠被仙台病毒，小鼠肝炎病毒，泰勒氏小鼠脑脊髓炎病毒/ GDVII株，小鼠微小病毒，小鼠巨细胞病毒，呼吸道肠道孤儿病毒（Reo-3病毒），小鼠细小病毒，小牛轮状病毒的抗原包被婴儿腹泻病毒，痘苗病毒痘苗病毒和肺炎支原体。通过接种BALB / cj和C57BL / 6j小鼠，产生了对单个传染原的抗体单阳性的标准血清。使用涂覆有上面列出的10种传染原的抗原的微珠混合物，对这些实验感染的小鼠的血清以及自然感染的小鼠的血清进行分析。结果表明，对于血清检测，多重测定至少与ELISA一样灵敏和特异性。重要的是，多重分析仅需1微升血清即可在一个反应​​容器中同时进行10种小鼠传染剂的血清学检测。因此，这种多重微珠测定是一种可靠，有效且具有成本效益的诊断方式，将影响研究中所用小鼠的血清监测。

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