Key Laboratory of Luminescence and Real-Time Analytical Chemistry (Ministry of Education), College of Pharmaceutical Sciences, Southwest University, Chongqing, 400716, China.
Key Laboratory of Luminescence and Real-Time Analytical Chemistry (Ministry of Education), College of Pharmaceutical Sciences, Southwest University, Chongqing, 400716, China.
Talanta. 2019 Dec 1;205:120130. doi: 10.1016/j.talanta.2019.120130. Epub 2019 Jul 9.
Bacterial analysis and antibiotic resistance testing (ART) are of great significance in clinical diagnosis and therapy of bacterial infectious diseases. In this work, a portable antibiotic-affinity chromatographic test strip has been developed for rapid analysis of Staphylococcus aureus (S. aureus) and further applied for ART of this pathogen. Porcine IgG was immobilized on a nitrocellulose membrane for capturing S. aureus based on the selective binding capability of the Fc fragment of IgG toward protein A on the surface of the target bacteria. Fluorescent microspheres modified with teicoplanin (TEI) were applied as signal substances to trace S. aureus utilizing the hydrogen bond conjugation between this antibiotic and Gram-positive bacteria. S. aureus can be analyzed within the concentration range from 1.4 × 10 CFU mL to 1.4 × 10 CFU mL. The recovery values for spiked samples were 93.3-110.0%. The obtained results of ART for penicillin, daptomycin, gentamicin, cefoxitin and clindamycin against S. aureus showed agreement with those of traditional broth dilution method. The procedures for bacterial analysis and ART can be accomplished within 20 and 110 min, respectively. The antibiotic-affinity chromatographic test strip showed great promise in point-of-care testing because of its ideal portability and rapidity.
细菌分析和抗生素耐药性检测(ART)在细菌性传染病的临床诊断和治疗中具有重要意义。在这项工作中,我们开发了一种便携式抗生素亲和色谱测试条,用于快速分析金黄色葡萄球菌(S. aureus),并进一步将其应用于该病原体的 ART。基于 IgG 的 Fc 片段对目标细菌表面蛋白 A 的选择性结合能力,将猪 IgG 固定在硝酸纤维素膜上用于捕获 S. aureus。用替考拉宁(TEI)修饰的荧光微球被用作信号物质,利用这种抗生素与革兰氏阳性菌之间的氢键结合来追踪 S. aureus。可以在 1.4×10 CFU mL 至 1.4×10 CFU mL 的浓度范围内分析 S. aureus。加标样品的回收率为 93.3-110.0%。用传统肉汤稀释法对青霉素、达托霉素、庆大霉素、头孢西丁和克林霉素对 S. aureus 的 ART 结果进行了验证,结果一致。细菌分析和 ART 的步骤分别可以在 20 分钟和 110 分钟内完成。由于其理想的便携性和快速性,抗生素亲和色谱测试条在即时检测方面具有很大的应用前景。