Ramezani Mohammad, Abnous Khalil, Taghdisi Seyed Mohammad
Pharmaceutical Research Center, Mashhad University of Medical Sciences, Mashhad, Iran.
Department of Pharmaceutical Biotechnology, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran.
Methods Mol Biol. 2017;1572:403-420. doi: 10.1007/978-1-4939-6911-1_26.
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.
血清和食品中抗生素残留的检测和定量需求巨大。我们已经开发了用于检测链霉素的适体传感器,采用电化学和光学方法。在第一种方法中,基于核酸外切酶I(Exo I)、适体互补链(CS)、适体(Apt)-CS共轭物的拱形结构和金电极的组合,开发了一种用于灵敏且选择性检测链霉素的电化学适体传感器。所设计的电化学适体传感器对链霉素表现出高选择性,检测限(LOD)低至11.4 nM。此外,所开发的电化学适体传感器成功用于检测牛奶和血清中的链霉素,其检测限分别为14.1和15.3 nM。在第二种方法中,基于水相金纳米颗粒(AuNPs)和双链DNA(dsDNA)设计了用于检测链霉素的荧光猝灭和比色适体传感器。在没有链霉素的情况下,适体/荧光素标记的互补链dsDNA是稳定的,导致AuNPs通过盐桥聚集,颜色从红色明显变为蓝色,并发出强烈的荧光。比色和荧光猝灭适体传感器对链霉素表现出优异的选择性,检测限分别低至73.1和47.6 nM。所展示的适体传感器成功用于检测牛奶和血清中的链霉素。对于血清,荧光猝灭和比色适体传感器的检测限分别确定为58.2和102.4 nM。对于牛奶,荧光猝灭和比色适体传感器的检测限分别计算为56.2和108.7 nM。