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A dual-label time-resolved fluorescence immunoassay for the simultaneous determination of ferritin and β2
Zhi Liu1, Jing Huang1, Rui-Ming Ou1
1Department of Hematology, Guangdong NO. 2 Provincial People's Hospital, Guangdong, Guangzhou, China.
Insights
A new dual-label immunoassay accurately detects ferritin and β2-microglobulin for early lymphocytic leukemia screening. This method offers high sensitivity and specificity for effective patient monitoring.
Area of Science:
- Biochemistry
- Immunology
- Clinical Diagnostics
Background:
- Lymphocytic leukemia is a primary malignant hematopoietic tissue tumor.
- Early detection and monitoring are crucial for effective management.
- Existing diagnostic methods may require improvement in sensitivity and specificity.
Purpose of the Study:
- To develop a dual-label time-resolved fluorescence immunoassay (TRFIA).
- To simultaneously detect ferritin (FER) and β2-microglobulin (β2-MG).
- To enable early screening and follow-up surveillance of lymphocytic leukemia.
Main Methods:
- Utilized sandwich immunoassay for FER and competitive immunoassay for β2-MG.
- Employed anti-FER and anti-β2-MG antibodies with time-resolved fluorometry (TRF).
- Evaluated performance using clinical blood samples and compared with commercial assays.
Main Results:
- Achieved high linear correlation coefficients (R² > 0.99) for both FER and β2-MG standard curves.
- Demonstrated high sensitivity (FER: 8 ng/mL, β2-MG: 1 ng/mL) and excellent average recovery rates.
- Showed high correlation with commercial assays (R² > 0.989).
Conclusions:
- The developed dual-label TRFIA is highly sensitive, specific, and accurate.
- It serves as an effective method for early screening of lymphocytic leukemia.
- The assay is suitable for follow-up surveillance of both acute and chronic lymphocytic leukemia.
Background:
Lymphocytic leukemia is a kind of primary malignant tumor of hematopoietic tissue. The aim was to establish a dual-label time-resolved fluorescence immunoassay (TRFIA) for the simultaneous determination of ferritin (FER) and β2 -microglobulin (β2 -MG) for the early screening and follow-up surveillance of lymphocytic leukemia.
Methods:
The sandwich immunoassay was used to detect the concentration of FER, and the competitive immunoassay was used to detect the concentration of β2 -MG in serum. FER in serum was captured by anti- FER antibody immobilized on microtiter wells, and then banded together with another anti- FER labeled with europium(III) Eu3+ chelate, followed by fluorescence measurement using time-resolved fluorometry (TRF). Sm3+ labeled β2 -MG and β2 -MG samples were added to compete with a certain amount of anti-β2 -MG antibody, followed by fluorescence measurement using TRF. The performance of this dual-label TRFIA was evaluated using the clinical blood and compared with the commercial assays.
Results:
The linear correlation coefficient (R2 ) of the FER and β2 -MG standard curves were 0.9914 and 0.9927, respectively. The sensitivity for FER detection was 8 ng/mL (dynamic range 0-1000 ng/mL), the average recovery was 100.51%; The sensitivity for β2 -MG detection was 1 ng/mL (dynamic range 0-1000 ng/mL), the average recovery was 101.02%. High correlation coefficients (R2 ) were obtained between the commercial assays (R2 =.9966 for FER, and R2 =.9897 for β2 -MG).
Conclusion:
The present dual-label TRFIA has high sensitivity, specificity, and accuracy in clinical sample analysis. It is an effective detection method for the early screening and follow-up surveillance of the acute and chronic lymphocytic leukemia.
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