Exonuclease III/Cas12a Cascade Amplification Strategy and Smartphone-Based Portable Fluorescence Detector to Repurpose the Commercial AFP Strip for the POCT of Multiple RNAs

Peng, Xin1,2; Mei, Xuecui1,2; Liu, Xueyan3; Zhang, Guanghui4; Li, Yingchun1,2

2024-07-01

10.1021/acs.analchem.4c02366

ANALYTICAL CHEMISTRY   影响因子和分区

0003-2700

1520-6882

Point of care testing (POCT) of nucleic acid (NA) contributes to the timely disease diagnosis, like bacteria and virus screening in households or resource-constrained areas, but its development has always been stagnant. Herein, we proposed an exonuclease III cascaded with CRISPR/Cas12a (Exo-III/Cas12a) amplification strategy and constructed a smartphone-based portable fluorescence detector (SPFD) to repurpose the commercial alpha-fetoprotein (AFP) strip for the ultrasensitive and hand-held detection of NA samples. In detail, the target-initiated-Exo-III/Cas12a strategy realizes the signal amplification and liberates AFP from magnetic beads through the trans-cleavages of activated Cas12a toward the AFP aptamer. After magnetic separation and migration, the fluorescence signals of the test (F- T) and control (F (C)) lines on the AFP strip were digitally output by the SPFD, and the F (T)/F C was employed for the quantitative analysis to minimize external disturbances and improve accuracy. We experimentally assessed the universe applicability of the proposed NA-POCT platform toward miRNA-155, 16S rRNA of Staphylococcus aureus, and ORF1a/b RNA of Covid-19 pseudovirus, achieving favorable detection limits of 42 aM, 18 CFU/mL, and 87 copies/mu L, respectively. Moreover, its simplicity, universality, and admirable detection performance demonstrate a great potential in the aspect of rapidly transforming the existing POCT devices for multiple new applications at the time of need.

SCI ; EI

英语

其他

Key Program of Natural Science Foundation of Shenzhen[JCYJ20220818102218039] ; National Natural Science Foundation of China[82373831] ; Shenzhen Key Laboratory of Prevention and Treatment of Severe Infections[ZDSYS20200811142804014] ; Shenzhen Key Medical Discipline Construction Fund[SZXK045] ; Guangdong Province Innovation Team Project for Universities[2023KCXTD049]

Chemistry

Chemistry, Analytical

WOS:001283231500001

AMER CHEMICAL SOC

CHEMISTRY

Web of Science

1.Harbin Inst Technol, Sch Sci, Shenzhen 518055, Peoples R China
2.Harbin Inst Technol, Sch Chem & Chem Engn, Harbin 150001, Peoples R China
3.Southern Univ Sci & Technol, Jinan Univ, Affiliated Hosp 1, Clin Med Coll 2,Shenzhen Peoples Hosp,Sch Med,Dept, Shenzhen 518020, Peoples R China
4.Shenzhen Hengsheng Hosp, Clin Lab, Shenzhen 518102, Peoples R China

Peng, Xin,Mei, Xuecui,Liu, Xueyan,et al. Exonuclease III/Cas12a Cascade Amplification Strategy and Smartphone-Based Portable Fluorescence Detector to Repurpose the Commercial AFP Strip for the POCT of Multiple RNAs[J]. ANALYTICAL CHEMISTRY,2024.

Peng, Xin,Mei, Xuecui,Liu, Xueyan,Zhang, Guanghui,&Li, Yingchun.(2024).Exonuclease III/Cas12a Cascade Amplification Strategy and Smartphone-Based Portable Fluorescence Detector to Repurpose the Commercial AFP Strip for the POCT of Multiple RNAs.ANALYTICAL CHEMISTRY.

Peng, Xin,et al."Exonuclease III/Cas12a Cascade Amplification Strategy and Smartphone-Based Portable Fluorescence Detector to Repurpose the Commercial AFP Strip for the POCT of Multiple RNAs".ANALYTICAL CHEMISTRY (2024).