Bulge-loop tuned entropy-driven catalytic reaction and tag-encoded barcodes for multiplexed mutation detection

作者全名："Luo, Wang; Wang, Ting; Weng, Zhi; Bai, Shulian; Zhang, Li; Wu, You; Yu, Hongyan; Bai, Dan; Wang, Ding; Chen, Huajian; Guo, Yongcan; Yang, Yujun; Li, Junjie; Zhang, Zhang; Xie, Guoming"

作者地址："[Luo, Wang; Wang, Ting; Weng, Zhi; Zhang, Li; Wu, You; Yu, Hongyan; Bai, Dan; Wang, Ding; Chen, Huajian; Yang, Yujun; Li, Junjie; Zhang, Zhang; Xie, Guoming] Chongqing Med Univ, Coll Lab Med, Chongqing Med Lab Microfluid & SPRi Engn Res Ctr, Key Lab Clin Lab Diagnost,Chinese Minist Educ, Chongqing 400016, Peoples R China; [Bai, Shulian] Chongqing Med Univ, Dept Lab, Univ Town Hosp, Chongqing 401331, Peoples R China; [Guo, Yongcan] Southwest Med Univ, Clin Lab, Tradit Chinese Med Hosp, Luzhou 646000, Peoples R China"

通信作者："Xie, GM (通讯作者)，Chongqing Med Univ, Coll Lab Med, Chongqing Med Lab Microfluid & SPRi Engn Res Ctr, Key Lab Clin Lab Diagnost,Chinese Minist Educ, Chongqing 400016, Peoples R China."

来源：SENSORS AND ACTUATORS B-CHEMICAL

ESI学科分类：CHEMISTRY

WOS号：WOS:000822675500003

JCR分区：Q1

影响因子：8.4

年份：2022

卷号：358

期号：　

开始页：　

结束页：　

文献类型：Article

关键词：DNA mutation; Nucleic acid hybridization; Entropy-driven catalytic reaction; Encoded barcode; Mutiplexed detection

摘要："Sensitive and specific detection of single nucleotide variants (SNVs) has great potential for disease diagnosis and prognosis. Hybridization probes based on DNA strand displacement have shown high SNV discrimination and programmability. However, it remains a challenge to balance the specificity and sensitivity of hybridization reactions due to the limited adjustability of probe sequences. Herein, we inserted bulge-loops in probes to fine-tune the thermodynamic changes of the entropy-driven catalytic reaction (EDC), significantly improving the selectivity (the DF of CAC>CGC mutation was promoted from 17.66832 +/- 2.25913-72.54 +/- 8.21) and responsiveness of SNV identification. In addition, oligonucleotides with chromophores (Tags) were used to impart microspheres with different fluorescence characteristics, to construct a set of convenient and flexible barcodes. Coupled with the bulge-loops tuned EDC probes and the Tag-encoded barcodes, the designed biosensor achieved higher specificity (DF=194.86 +/- 62.11) and lower signal leakage. Most importantly, it allowed simultaneous detection of low-abundance five-plexed EGFR mutations in the same system. This work provided insights into a rational design of DNA cascade reaction networks and advanced the construction of customizable multiplexed detection systems in the suspension array technology."

基金机构："National Natural Science Foundation of China [82172369, 81972025]; Natural Science Foundation of Chongqing, China [cstc2021jcyj-msxmX0326]; Outstanding Project of Chongqing Medical University [BJRC202024, BJRC202010]"

基金资助正文："This research work was financially supported by the National Natural Science Foundation of China (82172369, 81972025), the Natural Science Foundation of Chongqing, China (cstc2021jcyj-msxmX0326) and the Outstanding Project of Chongqing Medical University (BJRC202024, BJRC202010)."