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Fluorescent aptasensor based on the MNPs-CRISPR/Cas12a-TdT for the determination of nasopharyngeal carcinoma-derived exosomes

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Abstract

A fluorescence aptasensor based on taking the advantage of the combination of magnetic nanoparticles (MNPs), terminal deoxynucleotidyl transferase (TdT), and CRISPR/Cas12a was developed for the determination of nasopharyngeal carcinoma (NPC)-derived exosomes. The MNPs can eliminate background interference due to their magnetic separation capability. TdT can form an ultra-long polynucleotide tail which can bind with multiple crRNA, generating a signal amplification effect. The trans-cleavage activity of CRISPR/Cas12a can be specifically triggered via the crRNA binding with DNA, resulting in the bi-labeled DNA reporter with fluorophore and quencher being cleaved. The excitation wavelength of the fluorescence spectra was 490 nm. Fluorescence spectra with emission wavelengths ranging from 511 to 600 nm were collected. Under the optimization condition, the fabricated fluorescence aptasensor for NPC-derived exosome determination exhibited excellent sensitivity and specificity, with the linear range between 500 to 5 × 104 particles mL−1 and the limit of detection of 100 particles mL−1. It can be used for the determination of NPC-derived exosomes in clinical samples, which has a considerable clinical potential and prospect. 

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Funding

These works were supported by the National Natural Science Foundation of China (81803065), the Sanming Project of Medicine in Shenzhen (No. SZZYSM202106004), and the Science and Technology Planning Project of Shenzhen Municipality (JCYJ20210324143006017).

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Authors and Affiliations

  1. Department of Radiotherapy, Hubei Cancer Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430074, Hubei, China

    Peng Yi & Yajuan Zhou

  2. Department of Pharmacy, Zigong Third People’s Hospital, Zigong, 643020, Sichuan, China

    Dan Luo

  3. Department of Otorhinolaryngology Head and Neck Surgery, Shenzhen Fuyong People’s Hospital, Shenzhen, 518103, Guangdong, China

    Zhong Gao

  4. Shenzhen Baoan Authentic TCM Therapy Hospital, Shenzhen, 518101, Guangdong, China

    Qinhua Chen

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  1. Peng Yi
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  4. Qinhua Chen
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  5. Yajuan Zhou
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Correspondence to Qinhua Chen or Yajuan Zhou.

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Peng Yi and Dan Luo are the co-first authors.

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Yi, P., Luo, D., Gao, Z. et al. Fluorescent aptasensor based on the MNPs-CRISPR/Cas12a-TdT for the determination of nasopharyngeal carcinoma-derived exosomes. Microchim Acta 190, 74 (2023). https://doi.org/10.1007/s00604-023-05657-7

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