Visual Assay for Methicillin-Resistant Staphylococcus aureus Based on Rolling Circular Amplification Triggering G-Quadruplex/Hemin DNAzyme Proximity Assembly

作者全名："Xie, Xiaolin; Cheng, Xiaoxue; Dong, Juan; Li, Jia; Jiang, Lanxin; Yang, Tiantian; Liao, Bing; Ding, Shijia; Liu, Qingsong; Luo, Fukang; Cheng, Wei; Chen, Junman"

作者地址："[Xie, Xiaolin; Cheng, Xiaoxue; Dong, Juan; Li, Jia; Jiang, Lanxin; Yang, Tiantian; Cheng, Wei] Chongqing Med Univ, Ctr Clin Mol Med Detect, Affiliated Hosp 1, Chongqing 400016, Peoples R China; [Xie, Xiaolin; Liao, Bing; Luo, Fukang] Chongqing Ninth Peoples Hosp, Dept Clin Lab, Chongqing 400700, Peoples R China; [Ding, Shijia; Chen, Junman] Chongqing Med Univ, Minist Educ, Coll Lab Med, Key Lab Clin Lab Diagnost, Chongqing 400016, Peoples R China; [Liu, Qingsong] Women & Childrens Hosp Beibei Dist Chongqing, Dept Clin Lab, Chongqing 400700, Peoples R China"

通信作者："Cheng, W (通讯作者)，Chongqing Med Univ, Ctr Clin Mol Med Detect, Affiliated Hosp 1, Chongqing 400016, Peoples R China.; Luo, FK (通讯作者)，Chongqing Ninth Peoples Hosp, Dept Clin Lab, Chongqing 400700, Peoples R China.; Chen, JM (通讯作者)，Chongqing Med Univ, Minist Educ, Coll Lab Med, Key Lab Clin Lab Diagnost, Chongqing 400016, Peoples R China."

来源：ANALYTICAL CHEMISTRY

ESI学科分类：CHEMISTRY

WOS号：WOS:000935601800001

JCR分区：Q1

影响因子：7.4

年份：2023

卷号：95

期号：5

开始页：3098

结束页：3107

文献类型：Article

关键词：　

摘要："Nowadays, infections caused by methicillin-resistant Staphylococcus aureus (MRSA) have constituted a new challenge for anti-infective treatment. Precise identification and rapid clinical diagnostics of MRSA from other methicillin-sensitive strains entail assays with robust diagnostic efficiency and simple operation steps. Sensitive detection of MecA gene is promising to indicate MRSA infection, but it is challenged by the lack of isothermal and simple strategies. A visual assay based on isothermal rolling circular amplification and G-quadruplex/hemin (G4/hemin) DNAzyme proximity assembly was proposed for the immediate, efficient, and cost-effective detection of MecA in simple operation steps and in a single tube. The presence of MecA specifically drove the formation of circular templates, which further triggered isothermal amplification. The amplified product offered abundant binding sites for DNA-grafted hemin probes to form a novel proximity-assembled G4/hemin DNAzyme structure for colorimetric changing diagnosis. This tandem-repeated novel DNAzyme possessed higher catalytic activity and a lower background signal than traditional G4/hemin DNAzyme, ensuring sensitive discrimination of MRSA (limit of detection: 9.6 pM). Assay stability and antimatrix interference capability enable clinical application, which shows compared diagnostic ability with classic methods (100% sensitivity and 100% specificity) but possesses more simplified procedures and shorter turnaround time (< 6 h). This colorimetric strategy in a nonsite-specific and hypersensitive manner holds foreseeable prospects in clinical diagnostic and research applications."

基金机构："National Natural Science Foundation of China [81873980, 81873972]; Chongqing Science Fund for Distinguished Young Scholars [cstc2019jcyjjqX0028]; Training Program for Advanced Young Medical Personnel of Chongqing [2017HBRC003]; Special Funding for Postdoctoral Research Projects (Chongqing); Chongqing Postdoctoral Science Foundation [2021M700630]; China Postdoctoral Science Foundation; [cstc2021jcyj-bshX0153]"

基金资助正文："This work was funded by the National Natural Science Foundation of China (81873980 and 81873972), the Chongqing Science Fund for Distinguished Young Scholars (cstc2019jcyjjqX0028), the Training Program for Advanced Young Medical Personnel of Chongqing (2017HBRC003), Special Funding for Postdoctoral Research Projects (Chongqing), China Postdoctoral Science Foundation (2021M700630), and Chongqing Postdoctoral Science Foundation (cstc2021jcyj-bshX0153)."