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Sensitive immunosensing of the carcinoembryonic antigen utilizing aptamer-based in-situ formation of a redox-active heteropolyacid and rolling circle amplification

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Abstract

A highly sensitive electrochemical scheme for the detection of the cancer biomarker carcinoembryonic antigen (CEA) is reported. An aptamer-based formation of redox-active molybdophosphate is combined with rolling circle amplification (RCA). Gold nanoparticles (AuNPs) were utilized as supporting matrix for the aptamer and the primer to increase the primer/aptamer loading. The formation of a sandwich structure of anti-CEA/CEA/AuNP − aptamer-primer onto a glassy carbon electrode surface enabled an effective RCA reaction. Square wave voltammetry was applied to record the current signal at a peak potential of 0.18 V vs. Ag/AgCl. Compared to the immunosensor without RCA, the sensitivity of the immunosensor with RCA is increased by about a factor of 6. In the latter case, a wider linear range was obtained for the determination of CEA (from 0.5 pg mL−1 to 1 ng mL−1) with a detection limit as low as 0.1 pg mL−1. The practical applicability of the method was studied by analyzing CEA in human serum samples. The results were found to be in good agreement with those obtained by a reference method. Conceivably, the method can be applied to other DNA amplifications and therefore has the potential of finding wide applications.

Schematic of an electrochemical aptamer based method for the detection of the cancer biomarker carcinoembryonic antigen (CEA). An aptamer generated electrochemical current technique is coupled to rolling circle amplification (RCA).

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Acknowledgments

The authors thank the support of this work by the National Natural Science Foundation of China (No. 21575165), Tianjin Clinical Research Center for Organ Transplantation Grant (No. 15ZXLCSY00070) and Science and technology fund of Tianjin Municipal Health Bureau (No. 2013 KZ034).

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Author notes

  1. Wentao Jiang and Lei Liu contributed equally to this work.

Authors and Affiliations

  1. Key Laboratory for Critical Care Medicine of the Ministry of Health, Tianjin Key Laboratory for Organ Transplantation, Department of Liver Transplantation, Tianjin First Center Hospital, Tianjin, 300192, China

    Wentao Jiang, Li Zhang, Qingjun Guo & Yujun Cui

  2. First Center Hospital Clinic Institute, Tianjin Medical University, Tianjin, 300192, China

    Lei Liu

  3. College of Chemistry and Chemical Engineering, Central South University, Changsha, 410083, China

    Minghui Yang

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  1. Wentao Jiang
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Correspondence to Wentao Jiang or Minghui Yang.

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Jiang, W., Liu, L., Zhang, L. et al. Sensitive immunosensing of the carcinoembryonic antigen utilizing aptamer-based in-situ formation of a redox-active heteropolyacid and rolling circle amplification. Microchim Acta 184, 4757–4763 (2017). https://doi.org/10.1007/s00604-017-2522-7

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  • DOI: https://doi.org/10.1007/s00604-017-2522-7

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