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Time-resolved Fluorescence Immunoassay (TRFIA) for the Simultaneous Detection of MMP-9 and Lp-PLA2 in Serum

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Abstract

Currently, atherosclerosis accounts for the majority of cardiovascular morbidity and mortality worldwide, and predicting the stability of atherosclerotic plaque is the main method to prevent atherosclerotic death. This study aims to establish a dual-label time-resolved fluorescence immunoassay (TRFIA) of matrix metalloprotein-9 (MMP-9) and lipoprotein-associated phospholipaseA2 (Lp-PLA2) to predict atherosclerotic plaque stability. A dual-label TRFIA was introduced for the simultaneous quantification of MMP-9 and Lp-PLA2 using fluorescent lanthanide (Eu3+ and Sm3+) chelates. The performance (sensitivity, specificity, accuracy, precision and reference intervals in different subjects) of this TRFIA was evaluated and compared with commercial kit. The sensitivity of the TRFIA for MMP-9 was 0.85 ng/mL and for Lp-PLA2 was 0.68 ng/mL with high affinity and specificity. The average recoveries were 94.58% to 109.82%, and 104.32% to 109.26%, respectively. All intra- and inter-assay CVs ranged from 3.10% to 5.46%. For the normal subjects, the cutoff value was 160.70 ng/mL for MMP-9 and 183.73 ng/mL for LP-PLA2; for the subjects with stable plaque, the cutoff value was 181.98~309.22 ng/mL for MMP-9 and 194.73~337.89 ng/mL for LP-PLA2; for the subjects with unstable plaque, the cutoff value was 330.43 ng/mL for MMP-9 and 343.23 ng/mL for LP-PLA2. This TRFIA detection results agreed well with the results of commercial kit (R2=0.9567 and R2=0.9771, respectively) in clinical serum samples. The TRFIA developed has a wide detection range and good sensitivity for the high-throughput simultaneous detection of MMP-9 and Lp-PLA2 in serum, which provides a new method for predicting the stability of atherosclerotic plaque.

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Data Availability

The data used and/or analysed in the current study are given in the submitted article.

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Funding

This study was supported by the Science and Technology Plan Development Project of Jilin Province (No. 20200201529JC).

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Authors and Affiliations

  1. Department of Cardiology, Beihua University Affiliated Hospital, Jilin, 132000, China

    Hong-yan Ma, Qian Mao, Yan-bin Zhu, Chun-li Cong, Shi-yu Zheng & Qi Zhang

  2. Guangzhou Youdi Biotechnology Co., Ltd, Guangzhou, 510663, China

    Cui-cui Chen & Lai-qing Li

Authors
  1. Hong-yan Ma
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  2. Qian Mao
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  3. Yan-bin Zhu
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  4. Chun-li Cong
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  5. Shi-yu Zheng
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  6. Qi Zhang
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  7. Cui-cui Chen
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  8. Lai-qing Li
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Contributions

Hong-yan Ma and Lai-qing Li: Conceptualization, Methodology and Writing- Original draft preparation. Qian Mao, Yan-bin Zhu, Chun-li Cong and Shi-yu Zheng: Serum samples collection. Qi Zhang: Samples screening. Cui-cui Chen: Method optimization. Qian Mao and Yan-bin Zhu: Software and Data analysis.

Corresponding author

Correspondence to Lai-qing Li.

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The Institutional Review Board of Beihua University Affiliated Hospital approved this study, and all subjects gave written informed consent.

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The authors declared that they have no conflicts of interest.

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Ma, Hy., Mao, Q., Zhu, Yb. et al. Time-resolved Fluorescence Immunoassay (TRFIA) for the Simultaneous Detection of MMP-9 and Lp-PLA2 in Serum. J Fluoresc 31, 1771–1777 (2021). https://doi.org/10.1007/s10895-021-02811-3

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  • DOI: https://doi.org/10.1007/s10895-021-02811-3

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