Kim Seon-Hyung, Yun Hyun-Young, Jeong Hee-Jin
Department of Biological and Chemical Engineering, Hongik University, 30016, Sejong, Republic of Korea.
Mol Biol Rep. 2023 May;50(5):4715-4721. doi: 10.1007/s11033-023-08342-3. Epub 2023 Mar 10.
Bursaphelenchus xylophilus is a pathogenic nematode that causes pine wilt disease (PWD). To prevent the rapid spread of this pathogen, developing a method for rapid and accurate detection of B. xylophilus is required.
In this study, we produced a B. xylophilus peroxiredoxin (BxPrx), which is a protein that is overexpressed in B. xylophilus. Using recombinant BxPrx as an antigen, we generated and selected a novel antibody that binds to BxPrx via phage display and biopanning. We subcloned the anti-BxPrx single-chain variable fragment-encoding phagemid DNA to mammalian expression vector. We transfected the plasmid into mammalian cells and produced a highly sensitive recombinant antibody that enabled nanogram order detection of BxPrx.
The sequence of anti-BxPrx antibody as well as the rapid immunoassay system described here can be applied for rapid and accurate diagnosis of PWD.
松材线虫是一种引起松树萎蔫病（PWD）的病原线虫。为防止这种病原体的快速传播，需要开发一种快速准确检测松材线虫的方法。
在本研究中，我们制备了一种松材线虫过氧化物还原蛋白（BxPrx），该蛋白在松材线虫中过表达。以重组BxPrx为抗原，通过噬菌体展示和生物淘选产生并筛选出一种与BxPrx结合的新型抗体。我们将编码抗BxPrx单链可变片段的噬菌粒DNA亚克隆到哺乳动物表达载体中。将该质粒转染到哺乳动物细胞中，产生了一种高灵敏度的重组抗体，能够以纳克级水平检测BxPrx。
本文所述的抗BxPrx抗体序列以及快速免疫分析系统可用于松树萎蔫病的快速准确诊断。
