4.5 Article

Establishment and clinical application of time-resolved immunofluorescence assay of lipoprotein-associated phospholipase A2

Journal

ANALYTICAL BIOCHEMISTRY
Volume 648, Issue -, Pages -

Publisher

ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ab.2022.114674

Keywords

Lipoprotein-associated phospholipase A2; Time-resolved fluorescence immunoassay; Breast cancer; Dyslipidemia

Funding

  1. Chinese National Natural Science Foundation [82172336]
  2. Social Development Fund of Zhejiang Province [LGF20H200008]
  3. Key Research and Development Project of Zhejiang Province [2022C03118]
  4. Key Research and Development Project of Xiaoshan District [2020207]

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The study established a highly sensitive method for the detection of Lp-PLA2 and found that serum Lp-PLA2 levels in breast cancer patients were significantly higher than in healthy subjects, suggesting a potential association between serum Lp-PLA2 and dyslipidemia in breast cancer.
Aim: This study aimed to establish a highly sensitive time-resolved fluorescence immunoassay (TRFIA) for the detection of serum lipoprotein-associated phospholipase A2 (Lp-PLA2) and evaluate the clinical application value of Lp-PLA2 in patients with breast cancer. Methods: The level of Lp-PLA2 was detected using the double-antibody sandwich method. First, the Lp-PLA2TRFIA method was established, and the method was evaluated on the basis of linearity, sensitivity, precision, specificity, and recovery rate. Then, the fluorescence counts in serum of healthy subjects and patients with breast cancer were detected by Lp-PLA2-TRFIA, and the levels of Lp-PLA2 were calculated using a standard curve. Results: Lp-PLA2-TRFIA had a wide linear range (43.48-2000 ng/mL). The intra-assay precisions of Lp-PLA2TRFIA ranged from 2.66% to 4.84% (<10%), and the inter-assay precisions were between 5.39% and 6.95% (<15%). No cross-reaction was observed among Lp-PLA2, Tumor-associated trypsinogen-2, and T-cell immunoglobulin mucin 3. In addition, the recovery rates were between 90% and 100%. The serum Lp-PLA2 levels of patients with breast cancer were significantly higher than those of healthy subjects. Conclusions: We successfully established a highly sensitive Lp-PLA2-TRFIA method, and found serum Lp-PLA2 may be associated with dyslipidemia in breast cancer and could be used for auxiliary diagnose.

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