Establishment of interleukin‐18 time‐resolved fluorescence immunoassay and its preliminary application in liver disease

Abstract

Background To establish a time‐resolved fluorescence immunoassay of interleukin (IL)‐18 (IL‐18‐TRFIA) and detect its concentration in different liver disease serum samples. Methods The IL‐18 coating antibody and the Eu3+‐labeled detection antibody were used for the IL‐18‐TRFIA to detect serum IL‐18 concentration in patients with liver cancer, hepatitis B, hepatitis C, autoimmune hepatitis, fatty liver disease, and healthy controls. The double‐antibody sandwich method was used and methodological evaluation was performed. Results The average intra‐ and inter‐assay coefficient of variation for IL‐18‐TRFIA was 4.80% and 5.90%, respectively. The average recovery rate was 106.19 ± 3.44%. The sensitivity (10.96 pg/mL) was higher than that obtained using the ELISA method (62.5 pg/mL). The detection range was 10.96–1000 pg/mL. IL‐6 and galectin‐3 did not cross‐react with IL‐18‐TRFIA. The serum concentration of IL‐18 was (776.99; 653.48–952.39 pg/mL) in hepatitis C, (911; 775.55–1130.03 pg/mL) in fatty liver, (1048.88; 730.04–1185.10 pg/mL) in liver cancer, and (949.12; 723.70–1160.28 pg/mL) in hepatitis B. Moreover, IL‐18 serum levels were significantly higher in patients than the healthy controls (483.09; 402.52–599.70/mL) (p < 0.0001). Autoimmune hepatitis with a serum IL‐18 concentration of 571.62; 502.47–730.31 pg/mL was not significantly different from the healthy controls (p > 0.05). Conclusion We established a highly sensitive IL‐18‐TRFIA method that successfully detected serum IL‐18 concentrations in different liver diseases. Furthermore, IL‐18 serum concentration was higher in patients with liver cancer, hepatitis C, hepatitis B, and fatty liver disease compared to healthy controls.

Establishment of a time‐resolved immunoassay for acute kidney injury based on the detection of Kim‐1

“…TRFIA has the many features, such as simple preparation, stable, and no radioactive contamination 24 . In addition, lanthanides have high specific activity as a marker, ideal stability, and minimal effect on biological activity 25 . The addition of the dissociation enhancement buffer in the measurement phase can increase the initial fluorescence intensity by a million times 26 …”

“…24 In addition, lanthanides have high specific activity as a marker, ideal stability, and minimal effect on biological activity. 25 The addition of the dissociation enhancement buffer in the measurement phase can increase the initial fluorescence intensity by a million times. 26 In this study, we established Kim-1-TRFIA for the first time to detect the concentration of Kim-1 in human serum.…”

Upregulated lncRNA HCG18 in Patients with Non-Alcoholic Fatty Liver Disease and Its Regulatory Effect on Insulin Resistance

“… 44 Zhang et al report that the serum expression of IL18 in patients with liver cancer and fatty liver disease is significantly elevated, suggesting IL18 may act as an activator of liver diseases. 45 Some limitations also exist in this study, for instance, the lack of liver tissue study and the NAFLD staging of experimental mice.…”