The Isolation, Passage And Application Of Chlamydia Pneumoniae

Objective:To isolate C.pneumonia from PBMC and conduct its culture,passage and proliferation.Method:Isolate PBMC from blood samples,and release the C.pneumonia from broken cells by 37%PEG,then centrifugate them with Hep-2 together.After 7-day's culture,these cells were broken with freeze-thaw cycle,and put into new Hep-2 cells,then centrifugate them together to finish the first passage of C.pneumonia.The second passage was conducted in the same way.At the same time,we used MIF and PCR methods to detect C.pneumonia in Hep-2 cells.Result:by the method of MIF,we found that Hep-2 cells fused with PBMC,Hep-2 cells of first passage,Hep-2 cells of second passage were all C.pneumonia positive.We also detected the existence of C.pneumonia DNA in Hep-2 cells of second passage with the method of PCR.Conclusion:This way can successfully achieve the isolation,culture and passage of C.pneumonia from PBMC. Objective:To evaluate the Cpn-Ag prepared in the first part on detecting Cpn-Ab.Methods:Observe the reaction between the self-made Cpn-Ag and Cpn-McAb from USA by Dot-ELISA detection.Then the Cpn-IgG,IgM,IgA in blood specimens of 268 patients were detected by MIF with self-made Cpn-Ag and Cpn-AR39-Ag from USA at the same time.Results:The result of Dot-ELISA showed the self-made Cpn-Ag reacted with McAb.In the detection of 268 serum samples,the accordance among the results of the self-made and AR-39 from USA was excellent for the IgM,IgG and IgA detection rate(＞90%).The self-made Cpn-Ag had good sensitivity and specificity.The positive LR＞10,30 for IgM,the negative LR＜0.1.By McNemar test,P＞0.05 showed that two antigens had no difference in detecting but a high concordance tie between them by Kappa 0.704.The differences in endpoint titers of three antibodies were of no major concern.Conclusion:The self-made Cpn-Ag has great specificity and sensitivity on detecting Cpn-Ab and high concordance tie with Cpn-AR-39-Ag from USA, it could replace Cpn-AR39-Ag to detect the clinical sera specimens for Cpn infection diagnoses.