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A microfluidic chip using Au@SiO2 array–based highly SERS-active substrates for ultrasensitive detection of dual cervical cancer–related biomarkers

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Abstract

In this work, a microfluidic chip using Au@SiO2 array–based highly active SERS substrates was developed for quantitative detection of squamous cell carcinoma antigen (SCCA) and carcinoembryonic antigen (CEA) associated with cervical cancer. The chip consisted of six functional units with pump-free design, enabling parallel detection of multiple samples in an automatic manner without external pumps and improving the portability. Ag nanocubes (AgNCs) were labeled with Raman reporters and coupled with antibodies (labeling) to prepare SERS tags, while the Au nanoparticle–modified SiO2 microsphere (Au@SiO2) array was conjugated with antibodies (coating) to generate the highly SERS-active capturing substrate. In the presence of target biomarkers, they were captured by SERS tags and capturing substrate, resulting in the formation of “sandwich” structures which were trapped in the detection chamber. As the immune reaction proceeded, a large number of “hot spots” were generated by the proximity of the Au@SiO2 array substrate and AgNCs, greatly amplifying SERS signals. With this chip, the limits of detection of the SCCA and CEA levels in human serum were estimated to be as low as 0.45 pg mL−1 and 0.36 pg mL−1, respectively. Furthermore, the good selectivity and reproducibility of this chip were confirmed. Finally, clinical serum samples were analyzed by this chip, and the outcomes were consistent with those of enzyme-linked immunosorbent assay (ELISA). Thus, the proposed microfluidic chip can be potentially applied for the clinical diagnosis of cervical cancer.

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Funding

This work was supported by the National Natural Science Foundation of China (No. 82072088), the Social Development Foundation of Jiangsu (No. BE2018684), Yangzhou Science and Technology Project (No. YZ2017075), Contract for Maternal and Child Health Research Projects in Jiangsu Province (No. F201809), Postgraduate Research & Practice Innovation Program of Jiangsu Province (No. SJCX22_1820), High-end talent support program of Yangzhou University, and the QingLan Project of Yangzhou University.

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Authors and Affiliations

  1. Institute of Translational Medicine, Medical College, Yangzhou University, Yangzhou, 225001, People’s Republic of China

    Yingyan Gu, Shengjie Ge, Yu Mao, Yuexing Gu,  Xiaowei Cao & Dan Lu

  2. Department of Obstetrics and Gynecology, College of Clinical Medicine, Yangzhou University, Yangzhou, 225001, People’s Republic of China

    Yingyan Gu, Zhiyue Li & Dan Lu

  3. Jiangsu Key Laboratory of Integrated Traditional Chinese and Western Medicine for Prevention and Treatment of Senile Diseases, Yangzhou University, Yangzhou, 225001, People’s Republic of China

    Yingyan Gu, Shengjie Ge, Yu Mao, Yuexing Gu,  Xiaowei Cao & Dan Lu

  4. The First Clinical College, Dalian Medical University, Dalian, 116011, People’s Republic of China

    Zhiyue Li & Dan Lu

  5. Jiangsu Key Laboratory of Experimental & Translational Noncoding RNA Research, Medical College, Yangzhou University, Yangzhou, 225001, People’s Republic of China

    Shengjie Ge, Yu Mao, Yuexing Gu &  Xiaowei Cao

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  1. Yingyan Gu
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Correspondence to Xiaowei Cao or Dan Lu.

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This work followed the ethical principles of medical research in the Helsinki declaration. All serum samples were collected with the approval of the Ethics Committee of Yangzhou University School of Clinical Medicine, China. And all donors signed an informed consent before serum collection.

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Gu, Y., Li, Z., Ge, S. et al. A microfluidic chip using Au@SiO2 array–based highly SERS-active substrates for ultrasensitive detection of dual cervical cancer–related biomarkers. Anal Bioanal Chem 414, 7659–7673 (2022). https://doi.org/10.1007/s00216-022-04296-w

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