[Development and effectiveness evaluation of time-resolved fluoroimmunoassay kit for detection of ].

Objective: To develop a kit of time-resolved fluoroimmunoassay (TRFIA) for detection of protein SjP38, and evaluate its effectiveness.

Methods: The anti 9G7 SjP38 monoclonal antibody was used as the capture anti-body coated with 96-hole plate, and the Eu3 labeled 1A6 monoclonal antibody was used as the detection antibody to establish the TRFIA SjP38 kit. In addition, the accuracy, sensitivity, precision, stability and coincidence rate to pathogenic diagnosis of the kit were evaluated.

Results: This established kit possessed high accuracy, wide linear range from 2 to 1 250 ng/ml, high sensitivity with the minimum detectable concentration of 0.14 ng/ml, and good precision (the coefficient variation of the intra- and inter-assay were 3.6% to 4.6% and 5.1% to 6.7%, respectively). The stability tests showed that the reagents could be stable for six months at 4 ℃, 7 d at 37 ℃. The positive and negative corresponding rates to the pathogen detection method were 95% and 100% respectively.

Conclusions: All the performance and detection indicators of the kit have reached the requirements of clinical test, but its clinical application still needs further validation.