Tandem blocking of PCR extension to form a single-stranded overhang for facile, visual, and ultrasensitive gene detection.

ABSTRACT: In order to detect a predetermined gene in a field test, a facile, visual, and ultrasensitive approach without the need of special and expensive machines is required. In this study, a gene in the Ebola virus was targeted as an example for diagnosis. The key strategy is to incorporate molecular blockers (azobenzene-bearing moieties or thymine dimers) in tandem in one of the PCR primers and stop the polymerase extension there to form a single-stranded overhang. The PCR product was added to the dispersion of gold nanoparticles which were labelled with a probe oligonucleotide. When the Ebola virus-specific gene existed in the specimen, the oligonucleotide on the gold particles formed a double-helix with the single-stranded overhang, and thus the dispersion remained red in color. In the absence

SUBMITTER: Sui Z 

PROVIDER: S-EPMC9080086 | biostudies-literature | 2018 Apr

REPOSITORIES: biostudies-literature