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SERS spectroscopy using Au-Ag nanoshuttles and hydrophobic paper-based Au nanoflower substrate for simultaneous detection of dual cervical cancer–associated serum biomarkers

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Abstract

Ultrasensitive detection of specific biomarkers in clinical serum is helpful for early diagnosis of cervical cancer. In this paper, a surface-enhanced Raman scattering (SERS)-based immunoassay was developed for the simultaneous determination of squamous cell carcinoma antigen (SCCA) and osteopontin (OPN) in cervical cancer serum. Au-Ag nanoshuttles (Au-AgNSs) as SERS tags and hydrophobic filter paper-based Au nanoflowers (AuNFs) as capture substrate were constructed into a sandwich structure which served as an ultrasensitive SERS-based immunoassay platform. Finite difference time domain simulation confirmed that the electromagnetic field coupled between the AuNFs had a prominent SERS signal enhancement effect, which improved the detection sensitivity. SERS mapping showed that hexadecenyl succinic anhydride hydrophobic treatment could prevent the analyte from being quickly absorbed by the filter paper and increase the retention time to be more evenly distributed on the filter paper substrate. The immunoassay platform was verified to have good selectivity and reproducibility. With this method, the detection limits of SCCA and OPN in human serum were as low as 8.628 pg/mL and 4.388 pg/mL, respectively. Finally, in order to verify the feasibility of its clinical application, the serum samples of healthy subjects; cervical intraepithelial neoplasia I (CINI), CINII, and CINIII; and cervical cancer patients were analyzed, and the reliability of the results was confirmed by enzyme-linked immunosorbent assay experiments. The constructed SERS-based immunoassay platform could be used as a clinical tool for early screening of cancers in the future.

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Funding

Financial support was from the National Natural Science Foundation of China (No. 81701825), the Social Development Foundation of Jiangsu (No. BE2018684), the Natural Science Foundation of the Jiangsu Higher Education Institutions of China (No.17KJB416012), Yangzhou Science and Technology Project (No.YZ2017075), and Contract for Maternal and Child Health Research Projects in Jiangsu Province (No. F201809).

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Authors and Affiliations

  1. Department of Obstetrics and Gynecology, College of Clinical Medicine, Yangzhou University, Yangzhou, China

    Dan Lu, Menglin Ran, Yifan Liu, Ji Xia & Xiaowei Cao

  2. The First Clinical College, Dalian Medical University, Dalian, China

    Dan Lu, Menglin Ran, Yifan Liu, Ji Xia & Xiaowei Cao

  3. Jiangsu Key Laboratory of Integrated Traditional Chinese and Western Medicine for Prevention and Treatment of Senile Diseases, Yangzhou University, Yangzhou, China

    Dan Lu & Xiaowei Cao

  4. Transformative Otology and Neuroscience Center, College of Special Education, Binzhou Medical University, Yantai, 264003, China

    Liyan Bi

  5. Institute of Translational Medicine, Medical College, Yangzhou University, Yangzhou, China

    Xiaowei Cao

  6. Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, College of Veterinary Medicine, Yangzhou University, Yangzhou, 225009, China

    Xiaowei Cao

Authors
  1. Dan Lu
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  2. Menglin Ran
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  3. Yifan Liu
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  4. Ji Xia
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  5. Liyan Bi
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  6. Xiaowei Cao
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Correspondence to Dan Lu or Xiaowei Cao.

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This study was approved by the Ethics Committee of the College of Clinical Medicine of Yangzhou University and followed the guidelines of the Declaration of Helsinki.

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Lu, D., Ran, M., Liu, Y. et al. SERS spectroscopy using Au-Ag nanoshuttles and hydrophobic paper-based Au nanoflower substrate for simultaneous detection of dual cervical cancer–associated serum biomarkers. Anal Bioanal Chem 412, 7099–7112 (2020). https://doi.org/10.1007/s00216-020-02843-x

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