The State Key Lab of Silkworm Geneome Biology, College of Pharmaceutical Sciences, Southwest University, Chongqing 400715, China.
College of Life Sciences, Southwest University, Chongqing 400715, China.
Anal Chem. 2023 Feb 21;95(7):3754-3760. doi: 10.1021/acs.analchem.2c04851. Epub 2023 Feb 9.
Non-tuberculosis mycobacteria (NTM) is one family of pathogens usually leading to nosocomial infections. Exploration of high-performance biological recognition agent plays a pivotal role for the development of point-of-care testing device and kit for detecting NTM. () is a NTM which has been frequently applied as an alternative model for highly pathogenic mycobacteria. Herein, a recombinant tail protein derived from mycobacteriophage SWU1 infecting was expressed with system and noted as GP89. It shows a fist-like structure according to the results of homology modeling and ab initio modeling. It is confirmed as a lipoarabinomannan (LAM) binding protein, which can recognize studied NTM genus since abundant LAM constructed with d-mannan and d-arabinan is distributed over the mycobacterial surface. Meanwhile an enhanced green fluorescent protein (eGFP)-fused GP89 protein was acquired with a fusion expression technique. Then GP89 and eGFP-fused GP89 were applied to establish a sensitive and rapid method for fluorescent detection of with a broad linear range of 1.0 × 10 to 1.0 × 10 CFU mL and a low detection limit of 69 CFU mL. Rapid and reliable testing of antimicrobial susceptibility was achieved by the GP89-based fluorescent method. The present work provides a promising recognition agent for studied NTM and opens an avenue for clinical diagnosis of NTM-induced infections.
非结核分枝杆菌（NTM）是一类病原体，通常导致医院获得性感染。探索高性能生物识别剂对于开发用于检测 NTM 的即时检测设备和试剂盒起着关键作用。（）是一种 NTM，常被用作高致病性分枝杆菌的替代模型。本文中，来源于感染的分枝杆菌噬菌体 SWU1 的重组尾部蛋白在系统中表达，并标记为 GP89。根据同源建模和从头建模的结果，它呈现出拳头状结构。它被证实是一种脂阿拉伯甘露聚糖（LAM）结合蛋白，可识别研究中的 NTM 属，因为大量由 d-甘露聚糖和 d-阿拉伯聚糖构成的 LAM 分布在分枝杆菌表面。同时，通过融合表达技术获得了融合有增强型绿色荧光蛋白（eGFP）的 GP89 蛋白。然后，将 GP89 和融合有 eGFP 的 GP89 用于建立一种用于检测的荧光检测方法，具有 1.0×10 到 1.0×10 CFU mL 的宽线性范围和 69 CFU mL 的低检测限。基于 GP89 的荧光方法可实现对抗菌药物敏感性的快速可靠检测。本工作为研究中的 NTM 提供了一种有前途的识别剂，并为 NTM 感染的临床诊断开辟了新途径。
