Departamento de Microbiologia, Imunologia e Parasitologia, Universidade Federal de Santa Catarina, UFSC, Florianópolis, Brazil.
Arch Virol. 2011 Jan;156(1):9-16. doi: 10.1007/s00705-010-0810-1. Epub 2010 Sep 29.
Despite the economic impact of the infectious myonecrosis virus (IMNV) on shrimp farms in several countries, no method for immunological detection is currently available. With the aim of developing immunodiagnostic methods for IMNV detection in infected shrimps, a recombinant fragment of the IMNV major capsid protein gene encoding amino acids 105-297 (rIMNV₁₀₅₋₂₉₇ was heterologously expressed in Escherichia coli and used to immunize Balb/c mice, generating monoclonal antibodies (MAbs). Six hybridomas were obtained, and four of these recognized the presence of IMNV in tissue homogenates from naturally infected shrimps by immunodot blot assay. Among these MAbs, three were able to detect a ~100-kDa protein, which corresponds to the predicted mass of the IMNV major capsid protein, as well as viral inclusion bodies in muscle fibroses by western blot and immunohistochemistry. Two MAbs showed high specificity and sensitivity, showing no cross-reaction with healthy shrimp tissues in any assays, indicating their usefulness for IMNV detection.
尽管传染性肌肉坏死病毒 (IMNV) 对多个国家的虾养殖场造成了经济影响，但目前尚无免疫检测方法。为了开发用于检测感染虾中 IMNV 的免疫诊断方法，本研究在大肠杆菌中异源表达了编码 IMNV 主要衣壳蛋白基因第 105-297 位氨基酸的重组片段（rIMNV₁₀₅₋₂₉₇），并用其免疫 Balb/c 小鼠，产生单克隆抗体（MAbs）。获得了 6 株杂交瘤，其中 4 株通过免疫斑点印迹分析识别出天然感染虾组织匀浆中的 IMNV。在这些 MAbs 中，有 3 株能够通过 Western blot 和免疫组织化学检测到约 100 kDa 的蛋白，这与 IMNV 主要衣壳蛋白的预测质量相对应，以及肌肉纤维化中的病毒包涵体。两种 MAbs 表现出高度的特异性和敏感性，在任何检测中均与健康虾组织无交叉反应，表明它们可用于 IMNV 检测。
