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Serodiagnosis of Mice Minute Virus and Mouse Parvovirus Infections in Mice by Enzyme-Linked Immunosorbent Assay with Baculovirus-Expressed Recombinant VP2 Proteins

机译:杆状病毒表达的重组VP2蛋白的酶联免疫吸附法对小鼠细小病毒和小鼠细小病毒感染的血清学诊断。

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Mice minute virus (MMV) and mouse parvovirus (MPV) type 1 are the two parvoviruses known to naturally infect laboratory mice and are among the most prevalent infectious agents found in contemporary laboratory mouse colonies. Serologic assays are commonly used to diagnose MMV and MPV infections in laboratory mice; however, highly accurate, high-throughput serologic assays for the detection of MMV- and MPV-infected mice are needed. To this end, the major capsid viral protein (VP2) genes of MMV and MPV were cloned and MMV recombinant VP2 (rVP2) and MPV rVP2 proteins were expressed by using a baculovirus system. MMV rVP2 and MPV rVP2 spontaneously formed virus-like particles that were morphologically similar to empty parvovirus capsids. These proteins were used as antigens in enzyme-linked immunosorbent assays (ELISAs) to detect anti-MMV or anti-MPV antibodies in the sera of infected mice. Sera from mice experimentally infected with MMV (n = 43) or MPV (n = 35) and sera from uninfected mice (n = 30) were used to evaluate the ELISAs. The MMV ELISA was 100% sensitive and 100% specific in detecting MMV-infected mice, and the MPV ELISA was 100% sensitive and 98.6% specific in detecting MPV-infected mice. Both assays outperformed a parvovirus ELISA that uses a recombinant nonstructural protein (NS1) of MMV as antigen. The MMV rVP2 and MPV rVP2 proteins provide a ready source of easily produced antigen, and the ELISAs developed provide highly accurate, high-throughput assays for the serodiagnosis of MMV and MPV infections in laboratory mice.
机译:小鼠微小病毒(MMV)和1型小鼠细小病毒(MPV)是已知能自然感染实验室小鼠的两种细小病毒,并且是当代实验室小鼠群体中发现的最普遍的传染原。血清学检测通常用于诊断实验室小鼠的MMV和MPV感染。但是,需要用于检测MMV和MPV感染小鼠的高精度,高通量血清学检测方法。为此,克隆了MMV和MPV的主要衣壳病毒蛋白(VP2)基因,并使用杆状病毒系统表达了MMV重组VP2(rVP2)和MPV rVP2蛋白。 MMV rVP2和MPV rVP2自发形成病毒样颗粒,其形态与空细小病毒衣壳相似。这些蛋白质在酶联免疫吸附测定(ELISA)中用作抗原,以检测感染小鼠血清中的抗MMV或抗MPV抗体。来自被MMV( n = 43)或MPV( n = 35)感染的小鼠的血清和未感染的小鼠( n = 30)的血清用于评估ELISA。 MMV ELISA对检测MMV感染的小鼠具有100%的敏感性和100%特异性,而MPV ELISA对检测MPV感染的小鼠具有100%的敏感性和98.6%特异性。两种检测均优于使用MMV的重组非结构蛋白(NS1)作为抗原的细小病毒ELISA。 MMV rVP2和MPV rVP2蛋白为容易产生的抗原提供了现成的来源,并且开发的ELISA为实验室小鼠MMV和MPV感染的血清学诊断提供了高精度,高通量的检测方法。

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