Photoluminescent Response of Poly(3-methylthiophene)-DNA Single Nanowire Correlating to Nucleotide-Mismatch Locus in DNA–DNA Hybridization

π-Conjugated polymers have become qualified candidates for biosensing owing to their unique optoelectronic properties and excellent biocompatibility. In this contribution, nucleotide mismatches in DNA hybridization, being variable in position, are reflected in a stark manner by poly(3-methylthiophene) (P3MT) nanowires (NWs), in which probe DNA sequence is properly functionalized. Selected as the systematic investigation are complementary target DNA (tDNA), random sequence DNA, and three kinds of 1-mer mismatched tDNAs with different mismatch loci away from the NW’s surface. Nanoscale optical observation of the single P3MT NWs in solid states reveals that the more distant the mismatch position is from the surface, the higher the photoluminescence (PL) occurs, while the complementary sequence yields the highest but the random one remains the lowest. Hence, the PL intensity increases with the relative length of the DNA–DNA hybridization from the surface. These results deliver a new basis that π-conjugated polymers can be potentially applicable to detailed nucleotide analyses as in single nucleotide polymorphism.