Fig 1.
Schematic illustration of the dual-labeling TRFIA for simultaneous detection of aCL IgG and IgM.
First, pipette 100μl/well pretreated standard substance or serum to microtiter plate that precoated with aCL antigen, and incubated the plate with shaking for 30 min at 25°C. A washing step removes unbound and unspecifically bound serum or plasma components. Second, added 100μl conjugate of buffer diluted Eu3+-labeled anti-human IgG and buffer diluted Sm3+-labeled anti-human IgM into each well, and incubated with shaking for 30min at 25°C. A second washing step removes unbound conjugate. Then, 200μl enhancement solution was pipette to each well and shaken for 5 min. Last, read the fluorescent intensity.
Fig 2.
The calibration curves for aCL IgG and IgM using proposed dual-labeling TRFIA.
Table 1.
Intra- and Inter-assay precision of dual-label assay for aCL IgM and aCL IgG in serum of controls.
It was analyzed by measuring three pools of mixed serum specimens with high, intermediate and low concentration of aCL IgM and IgG 25 times in one series (intra-assay) and in duplicate in ten different series.
Table 2.
Recovery rate of dual-label assay for aCL IgG.
Table 3.
Recovery rate of dual-label assay for aCL IgM.
Fig 3.
Correlation between ELISA and dual-label assay of aCL IgG (Fig 3a) and IgM (Fig 3b) in human sera (n = 52).
Fig 4.
The comparison of liner range between ELISA and dual-label TRFIA for the detection of aCL IgG (Fig 4a) and IgM (Fig 4b), respectively.
Table 4.
Clinical positive rate.
Table 5.
The strength and weakness of the dual-label TRFIA.