Journal
TALANTA
Volume 268, Issue -, Pages -Publisher
ELSEVIER
DOI: 10.1016/j.talanta.2023.125269
Keywords
Ligation; Transcription; Ramification; Pyrophosphate; Colorimetric detection; Isothermal
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In this study, a Pradeep Kumar (PK)-probe was combined with a ligation-transcription-ramified RCA (LTR) dual-amplification system for the isothermal colorimetric detection of miRNA 25-3P. The detection system showed extraordinary sensitivity and high specificity, enabling simple naked-eye qualitative detection of miRNA.
In this study, we combined a Pradeep Kumar (PK)-probe with a ligation-transcription-ramified RCA (LTR) dualamplification system for the isothermal colorimetric detection of miRNA 25-3P, where the PK-probe transformed from its pink color to colorless in the presence of the amplification byproduct pyrophosphate (PPi), thereby allowing the simple naked-eye qualitative detection of the miRNA. Through this double-amplification strategy, the limit of detection reached as low as 91.4 aM-quite extraordinary sensitivity for a colorimetric miRNA detection system based on absorbance readings. Our detection system also operated with high specificity, the result of using two different target-selective ligation steps (linear DNA ligation and circular DNA ligation) mediated by SplintR ligase, and so could discriminate single-mismatched from perfectly matched target sequences. We suspect that this ultrasensitive and selective PK-probe/LTR dual-amplification system should be a great colorimetric diagnostic for the detection of any miRNA with high efficiency.
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