Research Of AlphaLISA Method For Chlamydia Pneumoniae Detection

Based on the early epidemiological research on Chlamydia pneumoniae, this studyfurther deeply analyzes the distribution of antigen determinant in the majorouter-membrane protein of Chlamydia pneumoniae, prepares the immune animal model,multi-clone antibody and mono-clone antibody by expressing and purifying the specificrecombinant antigen through the gene engineering methods, establishes the highly-specificand highly-sensitive AlphaLISA assay method for Chlamydia pneumoniae, and preparesfor the late research on the standard determination kit.Chlamydia pneumoniae is one of12pathogenic microorganisms which may greatlyinfluence the health of Chinese people in the21stcentury. It can not only causepneumoniae, but also may be the important pathogen for various diseases (e.g. stroke,cardiovascular diseases, atherosclerosis and even senile dementia). Its infection does notcause the typical clinical symptoms, and its continuous potential infection may damage themultiple sites at multiple levels. In this study, the AlphaLISA assay method can moresensitively rapidly easily test the Chlamydia pneumoniae, and thus offer the basis formedication and treatment more in time.At present, Chlamydia pneumoniae can be tested through various methods, e.g.pathogen separation/culturing, immunohistochemical test, fluorescent antibody test andmicro-immunofluorescent test. However, these assay methods have a certain shortcomings,and can not rapidly accurately test the Chlamydia pneumoniae. Despite the highersensitivity and time-validity than the traditional assay methods, the microorganism-basednucleic acid diagnostic technique (e.g. PCR and fluorescent quantitative PCR) require thehigher investment and high cost.Homogeneous light initiated chemiluminescent immunoassay (AlphaLISA) is a newerELISA technique. At present, it has been widely applicable for clinical assay at abroad. Itpossesses various advantages, e.g. high sensitivity, free of washing, simple operation,strong anti-interference, flexible assay mode and low cost. It has already tended to furtherreplace the ordinary ELISA for antigen/antibody assay. In a word, in the future, the assay technique of Chlamydia pneumoniae will developinto fast assay, high sensitivity, high flux, high specificity, homogeneous phase, simpleoperation and accurate stable reliable assay results. By standing on the foothold, lookingon the future and holding the attitude of rigorous research, this study prepares the antigenthrough the molecular biological technique, prepares the antibody through the mono-clonalantibody technique, preliminarily establishes the AlphaLISA assay method by combiningwith the antigen-antibody reaction, offers the basis for further research on the Chlamydiapneumoniae, and lays a foundation for the future preparation of its assay kit.