The Preliminary Study Of A Rapid Detecting Technology For Staphylococcus Aureusenes Based On Immunobiosensor

Objective: This study preliminarily established a high specificity and sensitivity, rapid and low-cost detecting technology of Staphylococcus aureusenes by combining F0F1-ATPase and chromatophores. It is a new method for the infection clinic diagnosis in Obstetrics and Gynecology.Methods: A fluorescence indicator, F1300, can quantify the activity of F0F1-ATPase.Based on this,a complex was constructed which is Staphylococcus aureusenes antibody-biotin-streptavidin-biotin-βsubunit monoclonal antibody, and connected this complex toβsubunit of F0F1-ATPase within the chromatophore. F0F1-ATPase changed its rotation speed with the capturing of Staphylococcus aureusenes by the antibody anchored on the complex. The concentration of Staphylococcus aureusenes was measured according as the relative fluoresent value of F0F1-ATPase biosensor tagged by F1300.Results:1.The chromatophores mean diameter which withdraws approximately is 70nm. In its peroxide solution, symbolized that the peak is surveyed in 860 nm .2.F0F1-ATP enzyme which load on F1300-ch hydrolysis activeness is 42.56μmol/mgmin.3.The intensity change of F1300 fluorescence may reflect that the H+ transportation during the sequence of ATP combined.It is means closely related with F0F1-ATP enzyme's activeness.4.Theβsubunit in F0F1-ATP enzyme unifies the different load, Transport activity was measured according as the relative fluoresent value of F0F1-ATPase biosensor tagged by F1300.5.It creates a curve on the bacterial concentration and fluorescence value of the correlation. As the concentration increases, fluorescence declined. This method took on favorable gradient between 103 and 105 CFU/ml.6.This method only took 4.5h .Conclusion: In this study,we set up a new experimental system of fluorescence probe F1300-labeled F0F1-ATPase within chromatophores as an immunobiosensor and applied the fluorescence probe-biosensor in detection of Staphylococcus aureusenes.This new method is sensitive,low-cost and rapid.This research establish the quantity effect curve between vaccine density and fluorescence value .