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Immobilized rolling circle amplification on extended-gate field-effect transistors with integrated readout circuits for early detection of platelet-derived growth factor

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Abstract

Detection of tumor-related proteins with high specificity and sensitivity is important for early diagnosis and prognosis of cancers. While protein sensors based on antibodies are not easy to keep for a long time, aptamers (single-stranded DNA) are found to be a good alternative for recognizing tumor-related protein specifically. This study investigates the feasibility of employing aptamers to recognize the platelet-derived growth factor (PDGF) specifically and subsequently triggering rolling circle amplification (RCA) of DNAs on extended-gate field-effect transistors (EGFETs) to enhance the sensitivity. The EGFETs are fabricated by the standard CMOS technology and integrated with readout circuits monolithically. The monolithic integration not only avoids the wiring complexity for a large sensor array but also enhances the sensor reliability and facilitates massive production for commercialization. With the RCA primers immobilized on the sensory surface, the protein signal is amplified as the elongation of DNA, allowing the EGFET to achieve a sensitivity of 8.8 pM, more than three orders better than that achieved by conventional EGFETs. Moreover, the responses of EGFETs are able to indicate quantitatively the reaction rates of RCA, facilitating the estimation on the protein concentration. Our experimental results demonstrate that immobilized RCA on EGFETs is a useful, label-free method for early diagnosis of diseases related to low-concentrated tumor makers (e.g., PDGF) for serum sample, as well as for monitoring the synthesis of various DNA nanostructures in real time.

The tumor-related protein, PDGF, is detected by immobilizing rolling circle amplification on an EGFET with integrated readout circuit

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Notes

  1. The reference EGFET has its top layer metal in contact with the electrolyte directly, resulting in a large interfacial capacitance. Therefore, the coupling coefficient approximates 1.

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Acknowledgments

We gratefully thank Professor Chen and Dr. Chia-Wen Lo at the Department of Physical Medicine and Rehabilitation at the National Taiwan University Associate Hospital for providing the mouse serum sample. This research was financially supported by the Department of Health, Taiwan (DOH99-TD-N-111-003); National Science Council, Taiwan (98-2321-B-009-001); and Ministry of Science and Technology (103-2221-E-492-012, 103-2622-E-492-005-CC3, 104-2622-E-492-020-CC3, CZ-12-17-37-105). The chip fabrication is supported by the Chip Implementation Center, Taiwan.

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Authors and Affiliations

  1. Instrument Technology Research Center, National Applied Research Laboratories, Hsinchu, 300, Taiwan, Republic of China

    Ming-Yu Lin

  2. Institute of Electronics Engineering, National Tsing Hua University, Hsinchu, 300, Taiwan, Republic of China

    Wen-Yang Hsu, Yueh-Lin Chung & Hsin Chen

  3. Institute of Molecular Medicine and Bioengineering, National Chiao Tung University, Hsinchu, 300, Taiwan, Republic of China

    Yuh-Shyong Yang

  4. Department of Neurosurgery, Chang Gung Memorial Hospital, Chia-Yi, 61363, Taiwan, Republic of China

    Jo-Wen Huang

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Correspondence to Hsin Chen.

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Lin, MY., Hsu, WY., Yang, YS. et al. Immobilized rolling circle amplification on extended-gate field-effect transistors with integrated readout circuits for early detection of platelet-derived growth factor. Anal Bioanal Chem 408, 4785–4797 (2016). https://doi.org/10.1007/s00216-016-9568-y

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