An electrochemical biosensor for the detection of aflatoxin B1 based on the specific aptamer and HCR biological magnification.

Aflatoxin B1 (AFB1) is a highly toxic mycotoxin, which causes severe acute or cumulative poisoning. Therefore, it is important to develop sensitive and selective detection methods for AFB1 for the safety of food and medicinal herbs. Herein, we have developed a "signal-on" electrochemical aptasensor based on the high specificity of the aptamer and hybridization chain reaction (HCR) biological amplification for AFB1 detection. In this work, thiol-modified complementary DNA (cDNA) immobilized on the surface of a gold electrode (GE) served as an initiator DNA. When AFB1 was present, it competed with the cDNA for binding to the aptamers, which resulted in the detaching of aptamers from the cDNA-aptamer duplexes. Then, the single-stranded cDNA acted as an initiator to trigger the HCR signal amplification. Therefore, long double-stranded DNA (dsDNA) products were produced, which could load large amounts of methylene blue (MB) molecules to generate a distinct electrochemical signal. Under the optimized conditions, the proposed electrochemical aptasensor achieved the ultrasensitive detection of AFB1 with a linear detection range of 0.01-100 pg mL-1, and a limit of detection (LOD) down to 2.84 fg mL-1. Furthermore, the electrochemical aptasensor was successfully applied for detecting AFB1 in corn and two kinds of traditional Chinese medicine samples, indicating the potential value for AFB1 detection in practical samples.

A fluorescent aptasensor based on gold nanoparticles quenching the fluorescence of rhodamine B to detect acetamiprid.

Pesticide residue detection is one of the main safety issues in the utilization of medicinal plants. In this work, a highly selective and sensitive aptasensor for acetamiprid determination was designed. The mechanism of the proposed method is based on the fluorescence resonance energy transfer (FRET) between gold nanoparticles (AuNPs) and rhodamine B (RB). Aptamers protect AuNPs from salt-induced aggregation, which causes fluorescence quenching of RB by the AuNPs via surface energy transfer. In the absence of acetamiprid, AuNPs were coated with aptamers on the surface and dispersed in NaCl solution. At this time, the dispersed AuNPs could perfectly quench the fluorescence intensity of RB. In contrast, in the presence of acetamiprid, aptamers specifically combine with acetamiprid to form a complex. With a high salt concentration, AuNPs would be aggregated without aptamer protection, weakening the RB quenching effect. Therefore, the concentration of acetamiprid could be obtained from the change in fluorescence intensity in the system. A fluorescent sensing method was established with a linear range from 0.1 to 3 µg mL-1, and the LOD was 0.0285 µg mL-1. The recoveries of acetamiprid in traditional Chinese medicine (TCM) samples were 96.23-105.75%. This method has great application value for the detection of acetamiprid in a complex sample matrix.

Nanoparticle based bio-bar code technology for trace analysis of aflatoxin B1 in Chinese herbs.

A novel and sensitive assay for aflatoxin B1 (AFB1) detection has been developed by using bio-bar code assay (BCA). The method that relies on polyclonal antibodies encoded with DNA modified gold nanoparticle (NP) and monoclonal antibodies modified magnetic microparticle (MMP), and subsequent detection of amplified target in the form of bio-bar code using a fluorescent quantitative polymerase chain reaction (FQ-PCR) detection method. First, NP probes encoded with DNA that was unique to AFB1, MMP probes with monoclonal antibodies that bind AFB1 specifically were prepared. Then, the MMP-AFB1-NP sandwich compounds were acquired, dehybridization of the oligonucleotides on the nanoparticle surface allows the determination of the presence of AFB1 by identifying the oligonucleotide sequence released from the NP through FQ-PCR detection. The bio-bar code techniques system for detecting AFB1 was established, and the sensitivity limit was about 10-8 ng/mL, comparable ELISA assays for detecting the same target, it showed that we can detect AFB1 at low attomolar levels with the bio-bar-code amplification approach. This is also the first demonstration of a bio-bar code type assay for the detection of AFB1 in Chinese herbs.