Open AccessA dual‐label time‐resolved fluorescence immunoassay for the simultaneous determination of ferritin and β 2 ‐microglobulin
Author(s) -
Liu Zhi,
Huang Jing,
Ou Ruiming,
Yao Mengdong,
She Yanlin,
Chen Rui,
Li Cheng,
Xu Li,
Abudureyimu Aikebaier,
Zhang Qing,
Liu Shuang
Publication year - 2017
Publication title -
journal of clinical laboratory analysis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.536
H-Index - 50
eISSN - 1098-2825
pISSN - 0887-8013
DOI - 10.1002/jcla.22132
Subject(s) - chemistry , chromatography , fluorescence , immunoassay , beta 2 microglobulin , detection limit , ferritin , coefficient of variation , analytical chemistry (journal) , europium , antibody , biochemistry , immunology , medicine , ion , physics , organic chemistry , quantum mechanics
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 β 2 ‐microglobulin (β 2 ‐ 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 β 2 ‐ 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 ) Eu 3+ chelate, followed by fluorescence measurement using time‐resolved fluorometry ( TRF ). Sm 3+ labeled β 2 ‐ MG and β 2 ‐ MG samples were added to compete with a certain amount of anti‐β 2 ‐ 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 ( R 2 ) of the FER and β 2 ‐ 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 β 2 ‐ MG detection was 1 ng/mL (dynamic range 0‐1000 ng/mL), the average recovery was 101.02%. High correlation coefficients ( R 2 ) were obtained between the commercial assays ( R 2 =.9966 for FER , and R 2 =.9897 for β 2 ‐ 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.