Journal Article

Evaluation of a Chemiluminescent Enzyme Immunoassay for the Detection of Prostaglandin E‐Major Urinary Metabolite (PGE‐MUM)

Arihiro, Seiji, Saruta, Masayuki, Moriyama, Kazushige, Kato, Tomohiro, Sakurai, Toshiyuki, Wakabayashi, Mie, Katagiri, Noriko, Yagi, Shintaro, Nasu, Hidekazu, Takagi, Jun

2024

Overview

ABSTRACT Background We developed a fully automated quantitative immunoassay for the detection of prostaglandin E‐major urinary metabolite (PGE‐MUM). In this study, we evaluated the analytical performance of this assay. Methods Sensitivity, within‐run reproducibility, correlation with radioimmunoassay (RIA), cross‐reactivity, dilution linearity, spike recovery performance, analyte stability, and effects of coexisting substances were evaluated. The assay was also used to measure PGE‐MUM in 211 healthy people. Results The limit of detection and quantification were 1.0 and 1.3 ng/mL, respectively. When the assay was performed six times in a single run, the coefficient of variation ranged from 1.4% to 2.2%. The coefficient of correlation with a preceding RIA method was 0.970 with a correlation slope of 0.88. There was no cross‐reactivity with PGE‐MUM analogs. Linearity of dilution was confirmed at up to 16‐fold dilution with assay results within 100 ± 20% of the theoretical values calculated based on the undiluted sample. Spike recovery was good and ranged from 94% to 101%. Analyte stability was tested by storing samples at 25°C for 6 days, 10°C for 1 month, and by performing up to five freeze–thaw cycles. Assay results were all within 100 ± 10%, the values measured before storage and before the freeze–thaw process. Assay results in healthy people ranged from 3.1 to 162.7 ng/mL (mean: 35.8 ng/mL). After correction for creatinine, the 95% confidence interval was 8.68–42.25 μg/g creatinine. Conclusion The assay precisely detects PGE‐MUM. We developed a chemiluminescent enzyme immunoassay (CLEIA), which can detect prostaglandin E‐major urinary metabolite (PGE‐MUM) with automated procedure in ~30 min. Here, we evaluated its analytical performance. This assay can precisely detect PGE‐MUM with good reproducibility and showed a good correlation with preceding radioimmunoassay (RIA).

• Publisher: John Wiley & Sons, Inc,John Wiley and Sons Inc
• Subject: Acids / Adolescent / Adult / Aged / Antigens / Creatinine / Enzyme immunoassay / Enzymes / Female / Humans / Immunoassay / Immunoenzyme Techniques - methods / Inflammation / Informed consent / Limit of Detection / Luminescent Measurements - methods / Male / Middle Aged / PGE‐MUM / Polyclonal antibodies / Prostaglandin E / Prostaglandins E - metabolism / Prostaglandins E - urine / Radioimmunoassay / Radioimmunoassay - methods / Reproducibility / Reproducibility of Results / Sensitivity analysis / Urine / Young Adult