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. 2018 Mar 1;179:350-355. doi: 10.1016/j.talanta.2017.11.004. Epub 2017 Nov 6.
A novel dual-site recognition protocol was developed for chemiluminescent (CL) detection of Streptococcus mutans (S. mutans) based on a designed antibiotic-affinity strategy. Teicoplanin, a broad-spectrum antibiotic against Gram-positive bacteria, was adopted to functionalize magnetic particles and recognize S. mutans utilizing the strong affinity between this agent and D-Alanyl-D-Alanine peptide moieties in the bacterial cell wall. To achieve ideal specificity for S. mutans detection, rat immunoglobulin G2a (rat IgG2a) tagged with horseradish peroxidase (HRP) was used as the second recognition agent and signal tracer since Fab region of rat IgG2a could bind with streptococcal protein G highly expressed in the cell wall of S. mutans. Thus HRP-tagged sandwich complex of teicoplanin/S. mutans/rat IgG2a was formed on the magnetic particles, followed by a CL quantification of S. mutans based on a HRP-catalyzed luminol-HO-p-iodophenol CL reaction. This dual-site recognition protocol showed a linear range of 1.0 × 10-1.0 × 10 CFU mL and a detection limit of 33 CFU mL for S. mutans detection. The whole detection process could be completed within 70min. The recovery tests for food, environmental, pharmaceutical and biological samples showed acceptable recovery values between 83.0% and 110.0%, demonstrating its application potential for detection of bacteria in various sample matrixes.
基于设计的抗生素亲和策略，开发了一种用于化学发光（CL）检测变形链球菌（S. mutans）的新型双位点识别方案。替考拉宁是一种广谱抗生素，可用于对抗革兰氏阳性菌，被采用来功能化磁性颗粒，并利用该试剂与细菌细胞壁中 D-丙氨酰-D-丙氨酸肽部分之间的强亲和力来识别 S. mutans。为了实现对 S. mutans 检测的理想特异性，辣根过氧化物酶（HRP）标记的大鼠免疫球蛋白 G2a（rat IgG2a）被用作第二识别剂和信号示踪剂，因为 rat IgG2a 的 Fab 区域可以与高度表达在 S. mutans 细胞壁中的链球菌蛋白 G 结合。因此，HRP 标记的替考拉宁/S. mutans/rat IgG2a 夹心复合物在磁性颗粒上形成，随后基于 HRP 催化的鲁米诺-HO-p-碘苯酚 CL 反应对 S. mutans 进行 CL 定量。该双位点识别方案显示出 1.0×10-1.0×10 CFU mL 的线性范围和 33 CFU mL 的检测限，用于 S. mutans 检测。整个检测过程可在 70min 内完成。对食品、环境、药物和生物样品的回收测试显示出可接受的回收率在 83.0%至 110.0%之间，表明其在各种样品基质中检测细菌的应用潜力。
