Establishment Of The Quantitative Detection Method Of FluoroimmunochromatograpHy For TPSA And FPSA

Background Prostate specific antigen(Prostate specific antigen,PSA),secreted by the prostate epithelial tissue,molecular weight is 34 KD,It has been widely used in the diagnosis of prostate cancer,There are two types that can be detected by immunological methods,namely free prostate specific antigen(free-PSA,f PSA),prostate-specific antigen(PSA-ACT)that binds to ?1-antichrombin,commonly referred to as total prostate antigen(t PSA)is the sum of f PSA and PSA-ACT.It is deemed internationally that PSA level of less than 4ng / m L is normal,the risk of prostate cancer increases with increasing concentration of PSA,when PSA levels between 4-10 ng / m L is considered to be the diagnostic gray zone,t PSA and f PSA accounted t PSA percentage(% f PSA)are used to improve the diagnosis of prostate cancer diagnostic gray zone efficiency.Quantitative detection of t PSA and f PSA methods are mainly inclued that chemiluminescence method,fluorescence immunoassay time resolution method,electrochemical luminescence method and so on,these methods operate complicatly or need large instruments and equipment,and all of them are high throughput detection,they are suitable for the large and medium-sized hospitals,not suitable for determination of a single specimen,resulting in limiting the application of t PSA and f PSA testing in community hospitals.Currently,In the t PSA and f PSA detection kits,lack a detection method that can accurately and quickly quantify,operat simplly,The fluorescent microsp Here immune chromatograp Hy technology has the above advantages,it has promising to fill the blank of t PSA and f PSA detection method.Objective To set up a fluorescence immune chromatograp Hy assay for detection t PSA and f PSA,which is high sensitivity,strong specificity,excellent stability,good precision,also the linear range is wide.To supply a new convenient skill and method for dectecting t PSA and f PSA in clinic.Methods Use t PSA and f PSA to prepara indoor reference products.In this research,we use fluorescent microsp Heres conjugated to antibodies,We explored and promoted the experimental conditions in many ways using perfect test experimental technology and platform of fluoroimmunochromatograp Hy provided by Henan Biological Engineering Technology Research Center.We comprehensively evaluated the specificity,accuracy,linearity,precision and stability of the test strip.And compared the test method that our study established with Roche Kit(electrochemical luminescence method).Results The results show that the t PSA and f PSA indoor reference materials prepared in this experiment can be stored for at least 12 months in the environment of ?-20 ?,and can be kept at least 4 weeks at 2-8 ?.The strip that we made didn't have cross reaction with triglyceride and bilirubin,and also didn't have cross reaction with AFP?CEA?CA125.The intra variation coefficients were all less than 10%,and the inter variation coefficients were all less than 15%.Test strip at 2-8 ? storage conditions at least stable stored for 6 months,at 37 ? storage conditions could be stored for 10 days stable.We tested the concentration of t PSA and f PSA of 87 clinical specimen by using the test method established in our study,which have been determined out definite value by the America Roche Kits(electrochemical luminescence method).The correlation coefficient between the result of our strips and Roche company's result were 0.9727 and 0.9888 respectively,and the regression equation was Y=0.9372X+0.2626?Y=0.9981X-0.0537 respectively.The correlations between the two method were well.Conclusions(1)Preparation of stable t PSA,f PSA indoor reference products.(2)Initially established simultaneously detect t PSA and f PSA chromatography quantitative detection of fluorescence.(3)Prepared a good performance t PSA and f PSA quantitative fluorescence chromatography test strip.