Creation Of A Novel Method Of Double Mono-cloned Antibody For Detecting Circular Immune Complex Of Chlamydia Pneumoniae

Objective: to establish a sensitive and special method of double mono-cloned antibody for examining the circular immune complex (CIC) in blood, and to further probe the relation between CIC of chlamydia pneumoniae (Cpn) and the coronary heart disease.Methods: This experiment was consisted of three steps. 1. At the first step, we planned to detect the CpnCIC existing in the serum according to the classical indirect double mono-cloned method. Covering the special goat anti human IgG antibody in the plate under 4# for 12 hours. The samples were added to react with the above fixed IgG antibody, as followed the special mouse anti human CpnTW183IgG. The peroxidase labeled goat anti mouse IgG could combine with CpnlgG on CpnCIC if it existed. At last, the result was got through the reaction of substrate. 2. At the second step, according to the result of the first step, we tried to create a novel method. Covering the special mouse anti human CpnTW183IgG under 4癈 for 12 hours, after this step ,we added CpnCIC extracted from the blood of patients with coronary heart disease by 7%PEG6000. As the peroxidase labeled goat anti human IgG could combine with antibody in CIC, we can get the positive rate of experimental group through the reaction of substrate. 3. At the third step, according to the Gel electrophoresis DNA-binding assay, we changed it partly to detect CpnCIC through the retarding belt which was caused by being combined with CpnAb under 37# for 2 hours. It was supposed that the belt of CpnCIC would lost or become lighter.Results: 1. The results of all the samples at the first step were positive; 2. The positive rate of CpnCIC of the experimental group is higher than that of the control group (63.8%/40%), (x2=7.78, p<0.05). 3. The retarding belt abstracted from the mixture of CpnlgG and the positive sample which was certaintied by the second step appeared, and the belt of CpnCIC lost correspondingly.Conclusions: 1. The classical method of double mono-cloned method could not beused to detect CpnCIC in serum. 2. It was a sensitive and special method of the double mono-cloned antibody for detecting CpnCIC, which could be used for the samples in clinic, and there is a relation between CIC of chlamydia pneumoniae and the coronary heart disease. 3. The novel Gel shift assay could detect CpnCIC existing in the blood finely.