A new amplified π-shape electrochemical aptasensor for ultrasensitive detection of aflatoxin B1.

There is a prompt need for determination of aflatoxin B1 (AFB1) in food products to avoid distribution and consumption of contaminated food products. In this study, an accurate electrochemical sensing strategy was presented for detection of AFB1 based on aptamer (Apt)-complementary strands of aptamer (CSs) complex which forms a π-shape structure on the surface of electrode and exonuclease I (Exo I). The presence of π-shape structure as a double-layer physical barrier allowed detection of AFB1 with high sensitivity. In the absence of AFB1, the π-shape structure remained intact, so only a weak peak current was recorded. Upon the addition of AFB1, the π-shape structure was disassembled and a strong current was recorded following the addition of Exo I. Under optimal conditions, the electrochemical signals enhanced as AFB1 concentrations increased with a dynamic range of 7-500pg/mL and a limit of detection (LOD) of 2pg/mL. The developed aptasensor was also used to analyze AFB1 spiked human serum and grape juice samples and the recoveries were 95.4-108.1%.

A novel fluorescent aptasensor based on silica nanoparticles, PicoGreen and exonuclease III as a signal amplification method for ultrasensitive detection of myoglobin.

Measurement of myoglobin (Mb) in human blood serum is of great interest for quick diagnosis of acute myocardial infarction (AMI). In this study, a novel fluorescent aptasensor was designed for ultrasensitive and selective detection of Mb, based on target-induced high fluorescence intensity, complementary strand of aptamer (CS), PicoGreen (PG) dye, exonuclease III (Exo III) and silica nanoparticles coated with streptavidin (SNPs-Streptavidin). The developed aptasensor obtains characteristics of SNPs as enhancers of fluorescence intensity, Exo III as an enzyme which selectively digests the 3'-end of double-stranded DNA (dsDNA), PG as a fluorescent dye which could selectively bind to dsDNA and high selectivity and sensitivity of aptamer (Apt) toward its target. In the absence of Mb, no free CS remains in the environment of SNPs-Streptavidin, resulting in a weak fluorescence emission. In the present of Mb, dsDNA-modified SNPs-Streptavidin complex forms, leading to a very strong fluorescence emission. The developed fluorescent aptasensor exhibited high specificity toward Mb with a limit of detection (LOD) as low as 52 pM. In addition, the designed fluorescent aptasensor was efficiently used to detect Mb in human serum.

A novel electrochemical aptasensor based on arch-shape structure of aptamer-complimentary strand conjugate and exonuclease I for sensitive detection of streptomycin.

Detection and quantitation of antibiotic residues in blood serum and animal foodstuffs are of great significance. In this study, an electrochemical aptasensor was developed for sensitive and selective detection of streptomycin, based on exonuclease I (Exo I), complimentary strand of aptamer (CS), Arch-shape structure of aptamer (Apt)-CS conjugate and gold electrode. The designed aptasensor inherits characteristics of gold including large surface area and high electrochemical conductivity, as well as high sensitivity and selectivity of aptamer toward its target, property of Arch-shape structure of Apt-CS conjugate to act as a gate and barrier for the access of redox probe to the surface of electrode and the function of Exo I as an enzyme which selectively digests the 3'-end of single stranded DNA (ssDNA). In the absence of streptomycin the gate remains closed. Thus, the electrochemical signal is weak. Upon addition of streptomycin, the Apt leaves the CS and binds to streptomycin and the Arch-shape structure is disassembled. Then, Exo I addition leads to a strong electrochemical signal. The designed electrochemical aptasensor exhibited high selectivity toward streptomycin with a limit of detection (LOD) as low as 11.4nM. Moreover, the developed electrochemical aptasensor was successfully used to detect streptomycin in milk and serum with LODs of 14.1 and 15.3nM, respectively.

Targeted delivery of Epirubicin to cancer cells by PEGylated A10 aptamer.

Clinical administrations of anthracyclines are limited by cardiotoxicity and myelosuppression. Targeted delivery of anticancer agents is especially important in reducing their side effects. In this work, A10 (Apt), an aptamer for prostate-specific membrane anytigen (PSMA), was applied for targeted delivery of Epirubicin (Epi) to LNCaP cells (PSMA(+)). Flow cytometry analysis showed that PEG-Apt-Epi complex was internalized effectively to LNCaP cells (PSMA(+)), but not to PC3 cells (PSMA(-)). This fact was confirmed by less cytotoxicity of PEG-Apt-Epi complex in PC3 cells in comparison with Epi alone. No significant change in viability between Epi- and complex-treated LNCaP cells was observed. In conclusion, PEG-Apt-Epi complex is an efficient and simple system for specific delivery of drug to PSMA-expressing cell lines.