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Specific chemiluminescent protocol for dual-site recognition of Streptococcus mutans utilizing strong affinity between teicoplanin and Gram-positive bacteria
Talanta ( IF 5.6 ) Pub Date : 2017-11-06 , DOI: 10.1016/j.talanta.2017.11.004
Xiaoxiao Su , Mengyao Wang , Yue Wu , Yong He , Zhifeng Fu

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-H2O2-p-iodophenol CL reaction. This dual-site recognition protocol showed a linear range of 1.0 × 102–1.0 × 106 CFU mL−1 and a detection limit of 33 CFU mL−1 for S. mutans detection. The whole detection process could be completed within 70 min. 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.



中文翻译:

利用替考拉宁和革兰氏阳性细菌之间的强亲和力,对变形链球菌进行双位识别的特定化学发光方案

基于设计的抗生素亲和策略,开发了一种新颖的双位点识别方案,用于化学发光(CL)检测变形链球菌S. mutans)。Teicoplanin是一种针对革兰氏阳性细菌的广谱抗生素,被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识别变形链球菌。为了达到理想的变形链球菌检测特异性,将用辣根过氧化物酶(HRP)标记的大鼠免疫球蛋白G2a(大鼠IgG2a)用作第二种识别剂和信号示踪剂,因为大鼠IgG2a的Fab区可以与链球菌结合蛋白G在变形链球菌细胞壁中高表达。因此替考拉宁/ HRP的标记夹心复合变形链球菌上形成磁性颗粒,随后的量化CL /大鼠IgG2a变形链球菌根据HRP催化鲁米诺-H 2 ö 2 -p -iodophenol CL反应。该双位点识别方案显示,变形链球菌的线性范围为1.0×10 2 –1.0×10 6 CFU mL -1,检出限为33 CFU mL -1检测。整个检测过程可以在70分钟内完成。食品,环境,药物和生物样品的回收率测试显示可接受的回收值在83.0%至110.0%之间,这表明了其在各种样品基质中检测细菌的应用潜力。

更新日期:2017-11-06
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