A highly sensitive and versatile transcription immunoassay using a DNA-encoding tandem repetitive light-up aptamer

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Title
A highly sensitive and versatile transcription immunoassay using a DNA-encoding tandem repetitive light-up aptamer
Author(s)
Jieun Sim; M S Baek; K H Lee; D M Kim; Ju Young ByunYong Beom Shin
Bibliographic Citation
Talanta, vol. 224, pp. 121921-121921
Publication Year
2021
Abstract
Highly sensitive and accurate measurements of protein biomarkers are crucial for early diagnosis and disease monitoring. Here we report a versatile detection platform for sensitive detection of a protein biomarker using a tandem repeat Spinach aptamer DNA-based transcription immunoassay, which is a immunoassay combined with transcription-assisted Spinach RNA aptamer generation. We designed a DNA template encoding spa tandem repetitive Spinach sequence for enhanced generation of an RNA aptamer. The tandem repeated Spinach DNA template is consist of multiple monomeric units which is composed of T7 promoter, Spinach-2 and terminator. After in vitro transcription, the fluorescence signal from the 16R (nR, n = number of repeats) DNA template was enhanced up to ~ 15-fold compared to a single form (1R) DNA template. Using tandem repeat DNA, the proposed transcription immunoassay showed a limit of detection (LOD) of 37 aM, which is 103-fold lower than that of the conventional enzyme-linked immunosorbent assay (ELISA). The results demonstrate substantial promise for the ultrasensitive detection of various biological analytes using simple ELISA techniques. The high sensitivity and reliability of the proposed transcription immunoassay offer great promise for clinical assays.
Keyword
Transcription immunoassayELISALight-up RNA aptamerSpinach aptamerTandem repeats
ISSN
0039-9140
Publisher
Elsevier
DOI
http://dx.doi.org/10.1016/j.talanta.2020.121921
Type
Article
Appears in Collections:
Critical Diseases Diagnostics Convergence Research Center > 1. Journal Articles
Division of Research on National Challenges > Bionanotechnology Research Center > 1. Journal Articles
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