| Streptococcus pneumonia(S.pneumococcus)is the most frequency detection,as community acquired pneumonia or the pathogenic bacteria of lower respiratory tract adapting just-in-time inventory.It’ a kind of Gram-positive coccus.S.pneuiriococcus can cause high prevalence rate and death rate not only in China but also in any other countries of the world.S.pneumococcus infection symptom is not only cause high frequency,but also evolve to serious illness easily,so it is important for us to select a appropriate antibiotic at the very beginning.Besides,from the perspective of the principles of treating infectious disease,the more earlier we found out the pathogenic bacteria,the more exacter treating method we could choose.So it is closely related to reduce health care costs in the future and it is very important to prevent the emergence of drug resistance bacteria.It is known that the cell wall polysaccharides of S.pneumococcus polysaccharide(C-PS)is the common antigen of all S.pnezunococcus strains.Existed in all serotypes of S.pneumococcus cell walls.It provided a possibility method for testing all S.pneumococcus type strains.This thesis set up a method for rapid detection of streptococcus pneumoniae by testing human blood sample.This method has a advantage of good accuracy,high sensitivity,easy to operate and so on.And the result won’t be influenced by antimicrobial treatment.It does not only afford diagnostic basis quickly,but also guide people how to correctly use the antibacterial agents.It provided an auxiliary diagnosis of quickly and effectively way to diagnose S.pneumococcus.This study include involved,S.pneumococcus fermentation,S.pneumococcus of cell wall polysaccharides purified,and its activation,Conjugation of C-PS to carrier proteins CRM 197,combining product immune animals,purification of antibody,and using it to establish rapid detection method of pneumonia(ELISA).We can come to the conclusion through this study:(1)The influence of the reaction conditions of C-PS purification process(pH is 11.0±0.5,constant temperature bath time of the alkali treatment is 60 min)on C-PS allocated inventory;(2)This article compares CNBr and NalO4 activation methods,compared the two treating methods can be obtained that the activated polysaccharide with CNBr method produces a good result.(3)Single administration of C-PS could not induce high titer antibody in mice.However,the C-PS-CRM197 glyeoconjugate induced high titer antibody and immunological memory was observed.(4)This article establishes a competitive ELISA method to detect C-PS in the sample.And verified the detection line is 18.4 ng/mL,repeatability CV is 4.7%and the RSD of precision is 4%. |
