The biochemical characteristics of pine wood nematode (PWN)-secretory antigen 571 (PWN-SA571) identified from the PWN secretome and PWN-infected pine tree extracts (PWNIPT) were investigated. PWN-SA571 has homologs in various nematodes in diverse families and a signal peptide at the amino-terminal end. Since it was not possible to express the full-length PWN-SA571 protein, two peptides showing high antigenicity were synthesized and used as antigens for generating monoclonal antibodies (Mabs). Mab-secreting fusion hybridoma cell lines were selected based on immunoreactivity to free peptide and BSA-conjugated peptide (BSA pep) antigens by enzyme-linked immunosorbent assay for establishing Mab-secreting hybridoma lines. Ascites containing PWN-SA571-specific Mabs showed stronger immunoreactivity to PWNIPT than to healthy pine tree extracts. In addition, PWN-SA571-specific Mabs could recognize less than 20 ng of BSA pep in lateral flow assays. Furthermore, purified biotin-conjugated PWN-SA571-specific Mab IgG or IgM were able to detect BSA pep (<15 ng) and PWN extracts (<600 ng). Taken together, these results demonstrate that PWN-SA571-specific Mabs could detect PWN or PWNIPT extracts by ELISA, LFA, or dot blot analysis.

研究了从PWN分泌组和感染PWN的松树提取物（PWNIPT）中鉴定的松木线虫（PWN）分泌抗原571（PWN-SA571）的生化特性。PWN-SA571在不同家族的各种线虫中具有同源物，并且在氨基末端具有信号肽。由于不可能表达全长PWN-SA571蛋白，因此合成了两个显示出高抗原性的肽，并将其用作产生单克隆抗体（Mabs）的抗原。根据对游离肽和BSA结合肽（BSA pep）抗原的免疫反应性，通过酶联免疫吸附法选择单克隆抗体分泌型融合杂交瘤细胞系，以建立单克隆抗体分泌型杂交瘤细胞系。含有PWN-SA571特异性单克隆抗体的腹水对PWNIPT的免疫反应性强于对健康松树提取物的免疫反应性。此外，PWN-SA571特异性单克隆抗体在侧向流动检测中可识别不到20 ng的BSA pep。此外，纯化的结合生物素的PWN-SA571特异性Mab IgG或IgM能够检测BSA pep（<15 ng）和PWN提取物（<600 ng）。综上所述，这些结果表明，PWN-SA571特异性单克隆抗体可以通过ELISA，LFA或斑点印迹分析检测PWN或PWNIPT提取物。