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Development of a novel ssDNA aptamer targeting cardiac troponin I and its clinical applications

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Abstract

Cardiac troponin I (cTnI) is a specific biomarker of acute myocardial infarction (AMI). However, cTnI detection kits prepared with antibodies have many defects. Nucleic acid aptamers are sequences of single-strand DNA or RNA that can overcome the deficiency of antibodies. Herein, sandwich ELONA methods were established based on aptamers. Two selected ssDNA aptamers (Apt3 and Apt6) showed high binding affinity and sensibility (Apt3: Kd = 1.01 ± 0.07 nM, Apt6: k = 0.68 ± 0.05) and did not bind to the same domain of cTnI. Therefore, these two aptamers can be applied to the ELONA methods. The detection range of cTnI using the dual-aptamer sandwich ELONA method was 0.05–200 ng/mL, and the bioanalytical method verification results can meet the national standard of Chinese Pharmacopoeia (2020 Edition). There was no difference between results of the dual-aptamer sandwich ELONA method and the diagnostic results of serum obtained from 243 people (P = 0.39, P ˃ 0.05). The sensitivity and specificity of the ELONA with cTnI in serum were 96.46% and 93.85%, respectively. Compared with the FICA kit, which is clinically used, the consequences of ELONA method are closer to the diagnostic results. This study suggests that the aptamers Apt3 and Apt6 have high affinity and strong specificity and that the dual-aptamer sandwich ELONA method has a wide detection range and can be used to determine cTnI in serum, with potential applications in the diagnosis of AMIs.

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Data availability

The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.

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Funding

This work was supported by the Guangzhou Science and Technology Plan Project (No. 2019030100143) and the Guangzhou Science and Technology Major Project (No. 201604020166), National Natural Science Foundation of China (No.81971742).

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Authors and Affiliations

  1. Department of Pharmacy, Biomedicine Research Center, Key Laboratory for Major Obstetric Diseases of Guangdong Province, The Third Affiliated Hospital of Guangzhou Medical University, Guangzhou, Guangdong, China

    Yi Cen, Wenting Zhu, Zhongwen Yuan, Senling Feng, Xiaomin Liu, Yuting Li & Pengke Yan

  2. Tongpeng Zhongxu Pharmaceutical Technology Company, Guangzhou, Guangdong, China

    Zhongping Wang, Yiqing Chen & Caiyan Lin

  3. Department of Pharmacology, Guangzhou Medical University, Guangzhou, Guangdong, China

    Peixiong Ke

Authors
  1. Yi Cen
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  2. Zhongping Wang
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  3. Peixiong Ke
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  4. Wenting Zhu
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  5. Zhongwen Yuan
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  6. Senling Feng
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  8. Caiyan Lin
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  9. Xiaomin Liu
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  10. Yuting Li
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  11. Pengke Yan
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Contributions

Y. C. conceived the study and drafted the manuscript. All authors revised the manuscript. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Pengke Yan.

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We confirmed that all protocols on human samples were approved by the Medical Ethics Committee of the Third Affiliated Hospital of Guangzhou Medical University.

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Cen, Y., Wang, Z., Ke, P. et al. Development of a novel ssDNA aptamer targeting cardiac troponin I and its clinical applications. Anal Bioanal Chem 413, 7043–7053 (2021). https://doi.org/10.1007/s00216-021-03667-z

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  • DOI: https://doi.org/10.1007/s00216-021-03667-z

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