CPA-Cas12a-based lateral flow strip for portable assay of Methicillin-resistant <i>Staphylococcus aureus</i> in clinical sample

作者全名："Wu, Jiangling; Huang, Yu; Ding, Xiaojuan; Kang, Lina; Wang, Xiaoliang; Li, Dandan; Cheng, Wei; Liu, Gang; Xue, Jianjiang; Ding, Shijia"

作者地址："[Wu, Jiangling; Kang, Lina; Wang, Xiaoliang; Xue, Jianjiang] Chongqing Med Univ, Univ Town Hosp, Dept Clin Lab, Med Sci Res Ctr, Chongqing 401331, Peoples R China; [Huang, Yu] Chinese Acad Sci, Chongqing Inst Green & Intelligent Technol, Chongqing Key Lab Multiscale Mfg Technol, Chongqing 400714, Peoples R China; [Huang, Yu] Univ Chinese Acad Sci, Chongqing Sch, Chongqing 400714, Peoples R China; [Ding, Xiaojuan; Li, Dandan] Chongqing Med Univ, Affiliated Hosp 2, Dept Clin Lab, Chongqing 401331, Peoples R China; [Cheng, Wei] Chongqing Med Univ, Affiliated Hosp 1, Ctr Clin Mol Med Detect, Chongqing 400016, Peoples R China; [Ding, Xiaojuan; Li, Dandan; Liu, Gang] Chongqing Med Univ, Univ Town Hosp, Dept Crit Care Med, Chongqing 401331, Peoples R China; [Ding, Shijia] Chongqing Med Univ, Minist Educ, Coll Lab Med, Key Lab Clin Lab Diagnost, Chongqing 400016, Peoples R China"

通信作者："Xue, JJ (通讯作者)，Chongqing Med Univ, Univ Town Hosp, Dept Clin Lab, Med Sci Res Ctr, Chongqing 401331, Peoples R China.; Cheng, W (通讯作者)，Chongqing Med Univ, Affiliated Hosp 1, Ctr Clin Mol Med Detect, Chongqing 400016, Peoples R China.; Liu, G (通讯作者)，Chongqing Med Univ, Univ Town Hosp, Dept Crit Care Med, Chongqing 401331, Peoples R China."

来源：JOURNAL OF NANOBIOTECHNOLOGY

ESI学科分类：BIOLOGY & BIOCHEMISTRY

WOS号：WOS:001189209100001

JCR分区：Q1

影响因子：10.6

年份：2023

卷号：21

期号：1

开始页：　

结束页：　

文献类型：Article

关键词：Methicillin-resistant Staphylococcus aureus; Cross-priming amplification; CRISPR/Cas 12a; Lateral flow strip; Clinical sample; Clinical medication

摘要："The rapid and accurate identification of methicillin-resistant Staphylococcus aureus at an early antibiotic therapy stage would be benefit to disease diagnosis and antibiotic selection. Herein, we integrated cross-priming amplification (CPA) and CRISPR/Cas 12a (designated as CPA-Cas 12a) systems to establish a sensitive and efficient lateral flow assay to detect methicillin-resistant Staphylococcus aureus. This assay relies on the CPA isothermal nucleic acid amplification strategy which can amplify the DNA extracted from Staphylococcus aureus and accompanying the indiscriminately trans-cleavage process of Cas 12a/CrRNA duplex after recognizing specific sequence. Taking the advantage of reporter and high turnover Cas 12a activity, a dramatic change in response was achieved to produce a significant increase in the analytical sensitivity. The signal conversion and output were realized using a lateral flow strip to achieve field-deployable detection. Furthermore, this bioassay was accommodated with a microfluidic device to realize automatically portable detection. This proposed assay completed within 30 min with the detection limit of 5 CFU mL-1, was verified by testing bacterial suspension and 202 clinical samples. Given the high sensitivity, specificity and efficiency, this colorimetric readout assay through strip could be further promoted to the clinical diagnosis, clinical medication of multidrug-resistant bacteria."

基金机构："Project of Science and Technology Research Program of Chongqing Municipal Education Commission [KJQN202200431, KJQN202000444]; Natural Science Foundation of Chongqing [cstc2020jcyj-msxmX0190, cstc2019jcyj-msxmX0179, cstc2019jcyj-msxmX0294]; Project of the Basic and Frontier Funds in the Yuzhong District of Chongqing [20200154]; Talents Project of University-Town Hospital of Chongqing Medical University"

基金资助正文："This work was funded by the Project of Science and Technology Research Program of Chongqing Municipal Education Commission (Grant No. KJQN202200431 and KJQN202000444), the Natural Science Foundation of Chongqing (grant no. cstc2020jcyj-msxmX0190, cstc2019jcyj-msxmX0179 and cstc2019jcyj-msxmX0294), the Project of the Basic and Frontier Funds in the Yuzhong District of Chongqing (grant no. 20200154) and the Talents Project of University-Town Hospital of Chongqing Medical University."