Epitope Mapping of the Nucleocapsid Protein of Sendai Virus and Application of Antigenic Epitopes for the ELISA-Based Diagnosis of Sendai Virus Infection
Epitope Mapping of the Nucleocapsid Protein of Sendai Virus and Application of Antigenic Epitopes for the ELISA-Based Diagnosis of Sendai Virus Infection
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DOI：
10.1292/jvms.12-0496
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发表时间：
2013-07-01
影响因子：
1.2
通讯作者：
Agui, Takashi
中科院分区：
文献类型：
--
作者：
Asano, Atsushi;Torigoe, Daisuke;Agui, Takashi
仙台病毒（SeV）是感染实验室小鼠和大鼠的最普遍的病毒病原体之一。迄今为止，成熟的SeV病毒粒子已被用作血清学诊断的抗原。为了开发更特异和更容易准备用于诊断的抗原，我们用来自实验SeV感染小鼠的抗血清和包含覆盖整个NP的重叠10聚体肽的肽阵列膜检查了SeV的核衣壳蛋白（NP）中的抗原位点。我们在SeV-NP的氨基酸120-160和420-500这两个区域中发现了抗原性线性序列。从这些抗原性序列中，我们将分别对应于SeV-NP的位置119-134和458-474的两种合成肽IVKTRDMEYERTTEWL和FVTLHGAERLEEETNDE用作酶联免疫吸附测定（ELISA）中的抗原。使用这些肽的ELISA的评价揭示了它们对抗SeV抗血清是特异性的。此外，使用这些肽的ELISA能够在与商业ELISA试剂盒相同的程度上区分SeV阳性和SeV阴性小鼠血清。这些结果表明这些肽可用于SeV感染的血清学诊断。
Sendai virus (SeV) is one of the most prevalent viral pathogens infecting laboratory mice and rats. To date, mature SeV virions have been used as antigens for serological diagnosis. To develop antigens that are more specific and easier to prepare for diagnosis, we examined the antigenic sites in the nucleocapsid protein (NP) of SeV with antisera from experimentally SeV-infected mice and a peptide array membrane containing overlapping 10-mer peptides covering the entire NP. We found antigenic linear sequences in two regions, amino acids 120-160 and 420-500, of the SeV-NP. From these antigenic sequences, we applied two synthesized peptides, IVKTRDMEYERTTEWL and FVTLHGAERLEEETNDE, which correspond to positions 119-134 and 458-474 of the SeV-NP, respectively, as antigens in an enzyme-linked immunosorbent assay (ELISA). Evaluation of the ELISAs using these peptides revealed that they were specific to anti-SeV antisera. Furthermore, the ELISAs using these peptides were able to distinguish between SeV-positive and SeV-negative mouse sera to the same extent as a commercial ELISA kit. These results indicate that these peptides are useful for the serological diagnosis of SeV infection.