Biosensors and Biodetection

Mohammad Ramezani, Khalil Abnous, Seyed Mohammad Taghdisi

Ben Prickril, Avraham Rasooly

Detection and quantitation of antibiotic residues in blood serum and foodstuffs are in great demand. We have developed aptasensors for detection of streptomycin using electrochemical and optical methods. In the first method, an electrochemical aptasensor was developed for sensitive and selective detection of streptomycin, based on combination of exonuclease I (Exo I), complementary strand of aptamer (CS), arch shaped structure of aptamer (Apt)-CS conjugate, and gold electrode. The designed electrochemical aptasensor exhibited high selectivity toward streptomycin with a limit of detection (LOD) as low as 11.4 nM. Moreover, the developed electrochemical aptasensor was successfully used to detect streptomycin in milk and serum with LODs of 14.1 and 15.3 nM, respectively. In the second method, fluorescence quenching and colorimetric aptasensors were designed for detection of streptomycin based on aqueous gold nanoparticles (AuNPs) and double-stranded DNA (dsDNA). In the absence of streptomycin, aptamer/FAM-labeled complementary strand dsDNA is stable, resulting in the aggregation of AuNPs by salt bridge and an obvious color change from red to blue and strong emission of fluorescence. The colorimetric and fluorescence quenching aptasensors showed excellent selectivity toward streptomycin with limit of detections as low as 73.1 and 47.6 nM, respectively. The presented aptasensors were successfully used to detect streptomycin in milk and serum. For serum, LODs were determined to be 58.2 and 102.4 nM for fluorescence quenching and colorimetric aptasensors, respectively. For milk, LODs were calculated to be 56.2 and 108.7 nM for fluorescence quenching and colorimetric aptasensors, respectively.