Establishment Of Homogeneous Chemiluminescent Immunoassay For Serum Carbohydrate Antigen125

As one of the most common malignant cancer in women,ovarian cancer in new types of chemotherapy and targeted therapy drugs emerge in endlessly today its total mortality remains high; Its main reason is that ovarian cancer patients cannot receive early diagnosis, which makes early detection and diagnosis crucial. As a suggestion tumor maker with global agencies for risk factor screening, early diagnosis of genetic syndrome (individual) and differential diagnosis, curative effect and recurrence monitoring and prognosis judgement, carbohydrate antigen125(CA125) becomes very important.CA125is a antigen molecule detected by mouse monoclonal antibodies which product from ovarian serous adenoma cell line OVCA433immune mouse by Bast et,al. First CA125immunoassay method using OC125antibody to capture and detection at the same time and commercialization in1983.CA125measurements which present commonly used are enzyme-linked immunoassay, chemilu-minescence immunoassay and electrochemical luminescence immuneassay etc.Compared with traditional non-homogeneous immunoassay technology, the homogeneous luminescence immunoassay technology,which based on luminol oxygen channeling immunoassay of AlphaScreen platform,is high sensitivity, strong time resolution, wide linear range, low background fluorescence, disposable plates, stable, fast, simple, easy to miniaturization, automation, etc.Objective:Based on luminol oxygen channel immunoassay (LOCI) of AlphaScreen platform,we develope a homogeneous luminescence immunoassay system for carbohydrate antigen125(CA-125) detection and make a evaluation of methodology and clinical.Methods:Rabbit polyclonal antibody for CA125and mouse monoclonal antibody for CA125coated receptor microsbeads, biotin, respectively; composed of two antibodies, titration is used to determine the best combination;Titration method is used to determine the optimal concentration of the receptor microbeads and antibody; biotin and buffer of the detection system, reagent and sample order, and the proportion of reagent and sample and two phase reaction time were optimized. Based on optimizing the four parameter fitting equation of standard curve, according to the standard curve can be unknown concentration in clinical specimens by emitting signal value of CA125concentration. To detect the clinical specimens according to the curve and make methodology and clinical evaluation. Study by ELISA Calc software for statistical analysis.Results:This system used rabbit polyclonal antibody for CA125coated receptor microbeads, mouse monoclonal antibody for CA125and avidin coated donor microbeads for serum CA125, in appendix2manual test method as the best detection method;within the inner precision is3.93%-4.40%, between-run precision between5.60%-8.39%; Functional sensitivity of3.91U/ml, analysis sensitivity of3.49U/ml; Recovery was105.14%-105.14%; the interference rate of hemolysis, jaundice, and rheumatoid disease<10%; This paper established methods CA125reference limit26.53U/ml;50clinical specimen in the testing results and the Roche Elecsys2010electrochemiluminescence assay results with good correlation, r2=0.974; Z=0.623; the chi square test and consistency analysis indicate that the system and import electrochemiluminescence detection reagent are no statistical difference.Conclusion:This system can be used for quantitative detection of serum CA125, and with no washing, strong sensitivity, high accuracy, good repeatability, easy operation etc, and further research is expected to promote the clinical application.