Journal
ACS SENSORS
Volume 6, Issue 8, Pages 2928-2937Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acssensors.1c00695
Keywords
biosensor; AgNCs; fluorescence assay; BCR/ABL(p210) fusion gene; chronic myeloid leukemia
Funding
- National Natural Science Foundation of China [81572080, 81873972]
- Natural Science Foundation Project of Chongqing [cstc2015shmszx120086]
- Training Program for Advanced Young Medical Personnel of Chongqing [2017HBRC003]
- Chongqing Science Fund for Distinguished Young Scholars [cstc2019jcyjjqX0028]
- Foundation for Innovative Research Groups of Chongqing Higher Education Institutions [CXQT20013]
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A strategy based on nanocluster beacon (NCB) fluorescence was developed to identify the two isoforms of the BCR/ABL(p210) fusion gene in one-pot, which showed distinct differences in clinical manifestation, treatment effect, and prognosis risk. The fluorescence of AgNCs can be activated when they are placed in proximity to the corresponding enhancer sequences, allowing for successful identification of the two isoforms with high sensitivity and specificity. This strategy also realized isoform identification in leukemia cells and newly diagnosed CML patients within 40 minutes, providing a powerful tool to distinguish fusion gene subtypes simultaneously.
The BCR/ABL(p210) fusion gene is a classic biomarker of chronic myeloid leukemia, which can be divided into e13a2 and e14a2 isoforms according to different breakpoints. These two isoforms showed distinct differences in clinical manifestation, treatment effect, and prognosis risk. Herein, a strategy based on nanocluster beacon (NCB) fluorescence was developed to identify the e13a2 and e14a2 isoforms in one-pot. Because the fluorescence of AgNCs can be activated when they are placed in proximity to the corresponding enhancer sequences, thymine-rich (T-rich) or guanine-rich (G-rich). In this work, we explored an ideal DNA-AgNCs template as an excellent molecular reporter with a high signal-to-noise ratio. After recognition with the corresponding isoforms, the AgNCs can be pulled closer to the T-rich or G-rich sequences to form a three-way junction structure and generate fluorescence with corresponding wavelengths. Therefore, by distinguishing the corresponding wavelengths of AgNCs, we successfully identified two isoforms in one tube with the limitation of 16 pM for e13a2 and 9 pM for e14a2. Moreover, this strategy also realized isoform identification in leukemia cells and newly diagnosed CML patients within 40 min, which provides a powerful tool to distinguish fusion gene subtypes at the same time.
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