| 黄 茜,杨萍萍,张 强,廖雨晴,李晓彤.曲霉菌半乳甘露聚糖单克隆抗体的制备及初步应用[J].,2020,(7):1241-1245 |
| 曲霉菌半乳甘露聚糖单克隆抗体的制备及初步应用 |
| Preparation of Monoclonal Antibodies Against the Aspergillus Galactomannan and Its Preliminary Application |
| 投稿时间：2019-08-28 修订日期：2019-09-23 |
| DOI：10.13241/j.cnki.pmb.2020.07.008 |
| 中文关键词: 曲霉菌 半乳甘露聚糖 酶联免疫吸附法 单克隆抗体 |
| 英文关键词: Aspergillus Galactomannan ELISA Monoclonal antibodies |
| 基金项目:福建省教育厅中青年教师教育科研项目（JAT170013） |
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| 中文摘要: |
| 摘要 目的：制备抗曲霉菌半乳甘露聚糖的单克隆抗体，并基于获得的抗体建立用于快速准确检测曲霉菌感染的双抗体夹心酶联免疫吸附法（ELISA），以期可用于侵袭性曲霉菌病的临床诊断。方法：提取曲霉菌半乳甘露聚糖后免疫BALB/c小鼠，筛选与制备抗曲霉菌半乳甘露聚糖的单克隆抗体，通过间接ELISA法与Western Blot方法开展单克隆抗体检测性能分析，使用获得的单克隆抗体建立双抗体夹心ELISA方法，并初步应用于曲霉菌感染血清检测。结果：获得抗曲霉菌半乳甘露聚糖单克隆抗体5株，均可特异性识别曲霉菌半乳甘露聚糖，以其中性能最佳的3C9抗体和辣根过氧化物酶标记的3C9抗体配对为基础，建立了双抗体夹心ELISA方法，通过初步评价确定该方法可应用于临床侵袭性曲霉菌病血清检测，并且该方法与现有商品化试剂盒相比检测背景值较低，可更有效区分曲霉菌感染阴阳性血清。结论：本研究筛选获得针对曲霉菌半乳甘露聚糖的特异性单克隆抗体，以该抗体为基础建立双抗体夹心ELISA方法具有潜在转化应用前景，可为侵袭性曲霉菌病的临床诊断提供支持。 |
| 英文摘要: |
| ABSTRACT Objective: To prepare monoclonal antibodies (mAbs) against Aspergillus galactomannan (GM) and establish a rapid and accurate double-antibody sandwich ELISA for Aspergillus detection and for possible application in the diagnosis of invasive aspergillosis (IA). Methods: BALB/c mice were immunized with extracted GM from Aspergillus. MAbs against GM were screened and prepared. Indirect ELISA and Western Blot were used to evaluate the obtained mAbs. Then, a double-antibody sandwich ELISA for Aspergillus detection was established and preliminarily applied to detect Aspergillus infection using sera from patients. Results: Five strains of anti-GM monoclonal antibodies were obtained, all of which could specifically recognize the GM of Aspergillus. In this study, based on the best-performing mAb pairs of 3C9 and Horseradish Peroxidase (HRP)-conjugated 3C9, a double-antibody sandwich ELISA was established. Through preliminary evaluation, it was confirmed that this method could be applied to the clinical serum detection of IA. Compared with the existing commercial kits, this method could more effectively distinguish the negative and positive sera of Aspergillus infection with a much lower background. Conclusion: Specific mAbs against Aspergillus GM were obtained in this study and a double-antibody sandwich ELISA for Aspergillus detection was established, which has the potential for translation and could provide support for IA diagnosis. |
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