Journal
SENSORS AND ACTUATORS B-CHEMICAL
Volume 315, Issue -, Pages -Publisher
ELSEVIER SCIENCE SA
DOI: 10.1016/j.snb.2020.128049
Keywords
Aptamer; Isothermal amplification; Carbon nitride nanosheet; Time-resolved fluorescent nanoparticles; Aflatoxin
Funding
- National Natural Science Foundation of China [31871881]
- Zhangjiagang Science and Technology Support Plan (Social development) [ZKS 1803]
- Jiangsu Agriculture Science and Technology Innovation Fund (JASTIF) [CX (18) 2025]
- S&T Support Program of Jiangsu Province [BE2017623]
- National First-class Discipline Program of Food Science and Technology [JUFSTR20180303, JUSRP51714B]
- Distinguished Professor Program of Jiangsu Province
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Aflatoxin M-1 is a prevailing natural carcinogenic mycotoxin, found in milk. Herein, we reported a sensitive fluorescent aptasensor for AFM(1), based on rolling circle amplification (RCA) technology to improve the sensitivity of the assay. The detection of AFM(1) was achieved using KYF4: Eu3+ time-resolved fluorescent nanoparticles (KYF4: Eu3+ TRFNPs) as signal probe and graphitic carbon nitride (g-C3N4) nanosheet as a quencher. The AFM1 aptamer act as a recognition element and as a primer for DNA synthesis. TRFNPs attached to complementary DNA (TRFNPs-cDNA) acts as a signal probe. In the absence of target, a rolling circle template (RCT) ligated to aptamer and RCA reaction initiated. The TRFNPs-cDNA was strengthened to amplified RCA product (RCAP) to make RCAP/TRFNPs-cDNA duplex. This duplex cannot be adsorbed onto g-C3N4, and ultimately no fluorescence quenched. In the presence of AFM(1), aptamer recognized the target and aptamer-target complex was formed. Hence, no aptamer hybridized with RCT, therefore, no RCA and duplex formation. Consequently, free TRFNPs-cDNA adsorbed onto g-C3N4 and resulted in quenched fluorescence. The assay provides a lower detection limit (0.0194 pg/mL) than the previously reported assays and has good recoveries between 92-99.8 %. Hence, the present method would be a suitable method for the quantification of small molecules.
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