Detection of deletion mutations in DNA using water-soluble cationic fluorescent thiophene copolymer

• Published in: Talanta (Oxford) Vol. 79; no. 2; pp. 153 - 158
• Main Authors: Guan, Hongliang, Zhou, Peng, Zeng, Shang, Zhou, Xianglei, Wang, Yun, He, Zhike
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 15.07.2009; Elsevier
• Subjects: Analytical chemistry; Base Pairing; Cations; Chemical coupling; Chemistry; Conjugated copolymer; DNA - genetics; DNA Mutational Analysis - methods; Exact sciences and technology; Fluorescence; Hydrogen bond; Lifetime; Nucleic Acids - analysis; Polymers; Sequence Deletion; Solubility; Spectrometric and optical methods; Thiophenes; Fluorescence; Lifetime; Conjugated copolymer; Chemical coupling; Hydrogen bond; Water; Chemical bond; Doping; Thiophene; Aldehyde; Fluorescence spectrometry; DNA; Detection limit; Linearity; Copolymer; Energy transfer; Nucleic acid; Interaction energy
• ISSN: 0039-9140; 1873-3573
• DOI: 10.1016/j.talanta.2009.03.034

In this contribution, we designed a fluorescent thiophene copolymer to detect insertion/deletion mutation in DNA by doping aldehyde group in the main chain. The fluorescence of the copolymer could be dramatically quenched on the addition of single-stranded DNA (ssDNA) via strong electrostatic interactions and electronic/energy transfer. Although the complementary ssDNA made the fluorescence recover, the hydrogen bonds and chemical coupling also played a significant role between the unpaired bases and aldehyde group, which could differentiate the subtle differences in such mutant DNA. The influence of buffer pH, concentration of NaCl, heating time and the temperature was systemically investigated and the proposed method was then successfully applied to detect real sample. With the respect to the linearity, limit of detection precision, specificity, this procedure could provide sensitive methodologies for the rapid detection and identification of nucleic acids.