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Updated: Aug 6, 2026
Detection and Monitoring of Tumor Associated Circulating DNA in Patient Biofluids
Published on: June 8, 2019
Triple-amplified sequential activation allosteric DNA biosensor for in vivo quantitative detection of MYCN copy
Liang Zhao1, Yingyu Zhang2, Hengyun Tian3
1Health Commission of Henan Province Key Laboratory for Precision Diagnosis and Treatment of Pediatric Tumor, Henan Province Key Laboratory of Children's Genetic and Developmental Diseases, Henan International Joint Laboratory for Prevention and Treatment of Pediatric Disease, Children's Hospital Affiliated to Zhengzhou University Zhengzhou 450018 China zhangwancun@126.com houligong258@163.com +86-373-85515773.
Abstract:
Fluorescence in situ hybridization (FISH) is commonly used to detect the MYCN copy number (M/N ratio) in postoperative or biopsy-derived neuroblastoma (NB) tissue samples, a critical indicator for risk stratification of NB. However, most NB patients present with large tumors at an advanced stage, making tissue acquisition difficult both surgically and via biopsy. Therefore, a triple-amplified sequential activation allosteric DNA biosensor, termed TASA (comprising MYCN-TASA and NAGK-TASA), which leverages the elevated enzymatic activities of apurinic/apyrimidinic endonuclease 1 (APE1) and telomerase in the cytoplasm of NB cells as molecular switches and signal amplification molecules, was developed for in vivo quantitative detection of the M/N ratio of NB. MYCN-TASA and NAGK-TASA sequentially interact with APE1, MYCN (or NAGK), and telomerase, undergoing a conformational change that cyclically separates the fluorophore from the quencher with high efficiency, thereby producing a detectable fluorescence signal. MYCN-TASA and NAGK-TASA exhibit superior sensitivity, with limits of detection of 0.45 aM and 0.75 aM for MYCN mRNA and NAGK mRNA, respectively. In addition, MYCN-TASA and NAGK-TASA enable rapid in vivo quantitative determination of the M/N ratio in NB tissues. In particular, MYCN-TASA and NAGK-TASA enable accurate in situ quantification of the M/N ratio in clinical NB tissues. In summary, this study presents a novel strategy for quantifying the NB M/N ratio, demonstrating significant potential for future applications in biomedical research and clinical diagnosis.
