Multiplex Assay Kit for 1,25-Dihydroxyvitamin D3 (DHVD3) ,etc. by FLIA (Flow Luminescence Immunoassay)

This assay has high sensitivity and excellent specificity for detection of 1,25-Dihydroxyvitamin D3 (DHVD3) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between 1,25-Dihydroxyvitamin D3 (DHVD3) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Matrices listed below were spiked with certain level of 1,25-Dihydroxyvitamin D3 (DHVD3) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of 1,25-Dihydroxyvitamin D3 (DHVD3) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level 1,25-Dihydroxyvitamin D3 (DHVD3) ,etc. by FLIA (Flow Luminescence Immunoassay) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level 1,25-Dihydroxyvitamin D3 (DHVD3) ,etc. by FLIA (Flow Luminescence Immunoassay) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of 1,25-Dihydroxyvitamin D3 (DHVD3) ,etc. by FLIA (Flow Luminescence Immunoassay) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

1. Preparation of standards, reagents and samples before the experiment;
2. Add 50μL standard or sample to each well,
    add 10μL magnetic beads,and 50μL Detection Reagent A,incubate 60min at 37°C on shaker;
3. Wash plate on magnetic frame for three times;
4. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37°C on shaker;
5. Wash plate on magnetic frame for three times;
6. Add 100μL sheath solution, swirl for 2 minutes, read on the machine.