Bogale M, Sampson E, Hu W, Baniya A, Mishra S, Kwon K, Lopez-Nicora H D, DiGennaro P
University of Florida, Gainesville, FL 32611.
The Ohio State University, Columbus, OH 43210.
J Nematol. 2025 Jul 19;57(1):20250024. doi: 10.2478/jofnem-2025-0024. eCollection 2025 Feb.
Currently available nematode identification techniques rely on visual microscopic examination of their morphology and limited molecular assays. These methods generally serve their purpose of enumerating nematode genera and informing management recommendations. However, when identifying variations in pathogenicity or virulence within nematode populations and species - which is crucial for specific plant-parasitic nematode management recommendations - these methods are insufficient. Here, we demonstrate that nucleotide sequence information for tens of thousands of monoclonal antibodies (mAbs) can be generated for identification purposes using a single-cell RNA-seq of mature B cells obtained from mice immunized with nematode antigens. We also provide proof of concept by synthesizing two of these mAbs in vitro and demonstrate specificity using ELISA. Since mAbs can bind to a variety of molecules, their potential use may surpass discrimination among pathotype groups and shed light on what contributes to pathogenicity or virulence of nematodes that produce, or are associated with, these antigenic molecules.
目前可用的线虫鉴定技术依赖于对其形态的视觉显微镜检查和有限的分子检测。这些方法通常能实现对线虫属的计数并提供管理建议。然而，在识别线虫种群和物种内致病性或毒力的差异时（这对于特定植物寄生线虫的管理建议至关重要），这些方法并不充分。在此，我们证明，使用从用线虫抗原免疫的小鼠获得的成熟B细胞的单细胞RNA测序，可以生成数以万计单克隆抗体（mAb）的核苷酸序列信息用于鉴定目的。我们还通过体外合成其中两种mAb并使用ELISA证明其特异性，提供了概念验证。由于mAb可以与多种分子结合，它们的潜在用途可能超越致病型组之间的区分，并揭示产生或与这些抗原分子相关的线虫的致病性或毒力的影响因素。
