Analytical and Bioanalytical Chemistry

, Volume 408, Issue 17, pp 4785–4797 | Cite as

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

  • Ming-Yu Lin
  • Wen-Yang Hsu
  • Yuh-Shyong Yang
  • Jo-Wen Huang
  • Yueh-Lin Chung
  • Hsin Chen
Research Paper
First Online:
Received:
Revised:
Accepted:

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.

Graphical Abstract

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

Keywords

Real-time monitoring Label-free detection Rolling circle Amplification Extended-gate field-effect transistors Monolithically integrated circuits 
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Notes

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.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

216_2016_9568_MOESM1_ESM.pdf (187 kb)
ESM 1 (PDF 186 kb)

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Copyright information

© Springer-Verlag Berlin Heidelberg 2016

Authors and Affiliations

  • Ming-Yu Lin
    • 1
  • Wen-Yang Hsu
    • 2
  • Yuh-Shyong Yang
    • 3
  • Jo-Wen Huang
    • 4
  • Yueh-Lin Chung
    • 2
  • Hsin Chen
    • 2
  1. 1.Instrument Technology Research CenterNational Applied Research LaboratoriesHsinchuRepublic of China
  2. 2.Institute of Electronics EngineeringNational Tsing Hua UniversityHsinchuRepublic of China
  3. 3.Institute of Molecular Medicine and BioengineeringNational Chiao Tung UniversityHsinchuRepublic of China
  4. 4.Department of NeurosurgeryChang Gung Memorial HospitalChia-YiRepublic of China

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