A novel CRISPR/Cas14a-based electrochemical biosensor for ultrasensitive detection of Burkholderia pseudomallei with PtPd@PCN-224 nanoenzymes for signal amplification

作者全名："Li, Cai; Liu, Changjin; Liu, Rui; Wang, Yuexin; Li, Anyang; Tian, Shen; Cheng, Wei; Ding, Shijia; Li, Wenting; Zhao, Min; Xia, Qianfeng"

作者地址："[Li, Cai; Liu, Rui; Wang, Yuexin; Li, Anyang; Tian, Shen; Li, Wenting; Xia, Qianfeng] Hainan Med Univ, Dept Trop Dis, Sch Trop Med, Key Lab Trop Translat Med,Minist Educ,Affiliated H, Haikou 571199, Hainan, Peoples R China; [Liu, Changjin] Fifth Peoples Hosp Chongqing, Dept Lab Med, Chongqing 400062, Peoples R China; [Cheng, Wei; Ding, Shijia; Zhao, Min] Chongqing Med Univ, Coll Lab Med, Key Lab Clin Lab Diagnost, Minist Educ, Chongqing 400016, Peoples R China"

通信作者："Li, WT; Xia, QF (通讯作者)，Hainan Med Univ, Dept Trop Dis, Sch Trop Med, Key Lab Trop Translat Med,Minist Educ,Affiliated H, Haikou 571199, Hainan, Peoples R China.; Zhao, M (通讯作者)，Chongqing Med Univ, Coll Lab Med, Key Lab Clin Lab Diagnost, Minist Educ, Chongqing 400016, Peoples R China."

来源：BIOSENSORS & BIOELECTRONICS

ESI学科分类：CHEMISTRY

WOS号：WOS:000926856300001

JCR分区：Q1

影响因子：10.7

年份：2023

卷号：225

期号：　

开始页：　

结束页：　

文献类型：Article

关键词：CRISPR; Cas14a; PtPd@PCN-224; Nanoenzyme; Electrochemical biosensor; Burkholderia pseudomallei

摘要："In this work, PtPd nanoparticles functionalized porphyrin metal-organic framework nanoenzymes (PtPd@PCN224 nanoenzymes) are exploited as signal amplification tags to fabricate a rapid and ultrasensitive sensitive CRISPR/Cas14a-based electrochemical biosensor for Burkholderia pseudomallei (B. pseudomallei) specific DNA sequences detection. The prepared PtPd@PCN-224 nanoenzymes not only catalyze the reduction peak current of H2O2 to obtain a strong electrochemical signal output, but also provide massive active sites for the assembly of nucleic acids by Zr-O-P bonds. Besides, the designed target-activated CRISPR/Cas14a is able to recognize the target DNA sequences and further trigger the trans-cleavage of ssDNA for signal amplification. Benefiting from the target-activated CRISPR/Cas14a and PtPd@PCN-224 nanoenzymes, the developed electrochemical biosensor for B. pseudomallei DNA detection exhibits high sensitivity with detection of limit down to 12.8 aM and excellent specificity for distinguishing non-targeted bacteria. Moreover, the CRISPR/Cas14a-based electrochemical detection platform can also apply for other pathogenic bacteria diagnostic by well-designing sgRNA for target sequence recognition, possessing high flexibility and versatility in clinical diagnosis."

基金机构："Major Science and Technology Program of Hainan Province [ZDKJ202003, ZDKJ2021036]; National Natural Science Foundation of China [81960002, 82272431]; State Key Research Development Program of China [2022YFC2305000, 2022YFC2305003]; Hainan Province Clinical Medical Center, Science and Technology Plan of Hainan Province (Clinical Research Center)"

基金资助正文："This study was supported by the Major Science and Technology Program of Hainan Province (ZDKJ202003, ZDKJ2021036) , the Na- tional Natural Science Foundation of China (81960002, 82272431) , and State Key Research Development Program of China (2022YFC2305000, 2022YFC2305003) , and Hainan Province Clinical Medical Center, Sci- ence and Technology Plan of Hainan Province (Clinical Research Center) ."