Anti-Trinitrotoluene Aptamers: Design, Functional Assessment and Optimization

Abstract

The aim of this work is detection of trinitrotoluene (TNT) using aptamer as a new sensing strategy. Two pBluescript plasmids containing RT and ST anti-TNT aptamers were used as templates for aptamer amplification by PCR method. For this purpose, 126 bp ST-aptamer and 118 bp RT-aptamer were amplified using specific primers. TNT bound to bovine serum albumin (TNP-BSA) was used as the antigen, and digoxigenin (DIG)-labeled aptamers were detected by horseradish peroxidase conjugated to anti-DIG monoclonal antibodies. The sensitivity and specificity of ST- and RT-aptamers were determined using optimized PCR and enzyme-linked aptamer-sorbent assay. The sensitivity of this detection after optimization was determined about 1.76 nM of TNT using 1 pM of aptamers. These results indicated favorable functions of both aptamers for TNT detection that can be used in the future as aptasensors for investigation and utilization of the TNT identification.