Preparation Of The Dipstick Of Gold-immuno Chromayography Assay For The Diagnosis Of The Anbody Of Mycoplasma Pneumonia And Its Trial Check In Labortory

In order to diagnose the infection caused by Mycoplasma pneumoniae, a sensitive, specific, and rapid gold immunochromatogrophic assay (ICA) was developed, which is used to detect Mycoplasma pneumoniae antibodies in clinic human blood samples and is also suitable for grassroots institutes' use.Mycoplasma pneumoniae(Mp) is an important etiologic agent of tracheobronchitis and primary atypical pneumonia in children and adults. It is responsible for more than 20% cases of community-acquired pneumonia infection and becomes a major cause of severe pneumonia of those hospitalized aged patients.The clinical symptom of Mycoplasmal pneumonia is atypical. The infection of Mp is confirmed through serologic tests. The test of specific Mp-antibodies is still one of important methods, though some new clinical tests come into use recently, such as real-time PCR.Through sensitizing NC membrane with the purified and inactivated Mp proteins used as the antigen, reducing colloidal gold with trisodium citrate, labeling the Staphylococcus aureus protein (SPA) as the probe, the rapid gold-immunochromatographic agent detecting Mp antibodies was developed, following the theory of immuno-chromatograhy to diagnose Mycoplasma pneumonia. The presented study involves many parts, including preparation and identification of the complete inactivated Mp proteins, the optimization of the concentration and condition of antigen connecting, the optimization of the quantity of colloidal gold, the primary examination of the established immuo-colloidal gold method.As the result, Mp was successfully cultivated, purified and inactivated. The thalline grains were bright green, 0.3um in size, complete and round with clear boundary under the microscopes. According to the color reaction, the minimal concentrtion of stable SPA labeled by colloidal gold is 5μg per milliliter and the optimal one is 6ug SPA per milliliter.The NC membrane was sensitized with various concentrations of inactivated Mp by XYZ3000 spotting rite type BIO-DOT. The chromatic reaction results showed that the performance of the dipstick connected with 3mg/ml Mp antigen is the best one. So this concentration was chosen to sensitize NC membrane.In...