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Multiplex ratiometric gold nanoprobes based on surface-enhanced Raman scattering enable accurate molecular detection and imaging of bladder cancer

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Abstract

Recently, surface-enhanced Raman scattering (SERS) has been successfully used in the non-invasive detection of bladder tumor (BCa). The internal standard method was considered as an effective ratiometric strategy for calibrating signal fluctuation originated from the interference of measurement conditions and samples. However, it is still difficult to detect the target mRNA quantitatively using the current ratiometric SERS nanosensors. In this study, we developed an internal reference based ratiometric SERS assay. Two kinds of molecular beacons (MB) carrying Raman reporter molecules were anchored on sea-urchin-like Au nanoclusters (AuNCs). Thymidine kinase1 (TK1) MBs with hexachlorofluorescein (HEX) were used to capture tumor biomarker TK1 mRNA, and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) MBs with 5(6)-carboxyfluorescein (FAM) were used to offer internal standard signals. The internal reference GAPDH MB can reflect the consistent content of the GAPDH mRNA in single cells. The ratiometric method (I745/I645) can more accurately reflect the content of target mRNA in single cells. The ratiometric nanoprobes had excellent stability (coefficient of variation: 0.3%), high sensitivity (detection limit: 3.4 pM), high specificity (capable of single-base mismatch recognition) and ribozyme-resistant stability. Notably, the nanoprobes can effectively distinguish BCa cells from normal cells, and it was easy to contour the single BCa cell using the ratiometric method. By combining asymmetric polymerase chain reaction (PCR) and ratiometric nanoprobes, it was easy to distinguish the SERS ratio (I745/I645) as low concentration as 10−14 M. Further clinical detection in urine samples from patients with BCa confirmed its potential for early noninvasive diagnosis of BCa with the sensitivity of 80% and specificity of 100%, which is superior to the current urine cytological method.

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (No. 81901838), the Key Research and Development Plan in Shaanxi Province (Nos. 2020SF-123 and 2020SF-195), the Natural Science Foundation of Zhejiang Province (No. LQ21H160041), and the Medical Research Program of Department of Science and Technology of Xi’an, Shaanxi Province (No. 2019115713YX012SF048(4)). We thank Mengzhao Zhang, Qiuya Shao, Lu Wang and Lu Zhang at Department of Urology, the First Affiliated Hospital, Xi’an Jiaotong University for their support. We also thank Dr. Yu Wang at Instrument Analysis Center of Xi’an Jiaotong University for her assistance.

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  1. Xiao Liang, Pu Zhang, and Minghai Ma contributed equally to this work.

Authors and Affiliations

  1. Department of Urology, The First Affiliated Hospital, Xi’an Jiaotong University, Xi’an, 710061, China

    Xiao Liang, Pu Zhang, Minghai Ma, Tao Yang, Rui Zhang, Minxuan Jing, Rundong Song & Jinhai Fan

  2. Department of Thoracic Surgery, Tangdu Hospital, Air Force Medical University, Xi’an, 710038, China

    Xiao Liang & Lei Wang

  3. Department of Urology, Zhejiang Provincial People’s Hospital, People’s Hospital of Hangzhou Medical College, Hangzhou, 310014, China

    Pu Zhang

  4. State Key Laboratory of Bioelectronics, School of Biological Science and Medical Engineering, National Demonstration Center for Experimental Biomedical Engineering Education, Southeast University, Nanjing, 210096, China

    Xiangwei Zhao

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  1. Xiao Liang
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  2. Pu Zhang
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  4. Tao Yang
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Correspondence to Lei Wang or Jinhai Fan.

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Multiplex ratiometric gold nanoprobes based on surface-enhanced Raman scattering enable accurate molecular detection and imaging of bladder cancer

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Liang, X., Zhang, P., Ma, M. et al. Multiplex ratiometric gold nanoprobes based on surface-enhanced Raman scattering enable accurate molecular detection and imaging of bladder cancer. Nano Res. 15, 3487–3495 (2022). https://doi.org/10.1007/s12274-021-3902-1

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