The Establishment Of LiCA-MPO Assay And The Primary Application In The Coronary Artery Disease

Coronary artery disease is an important cause of death for adult in our contry. In developed countries, it is the first cause of death. Although the incidence and mortality of coronary artery disease is lower in our country than in developed countries,with the development of society and economy,it is increasing year by year. MPO is not only the media of oxidative stress but also the marker of systematic inflammation. it promotes the formation of the atherosclerosis by oxidative modification and plays an improtant role in the process of developement and evolvement of atherosclerosis plaque.Percutaneous coronary interventions(PCI)is an effective treatment to Coronary artery disease,but restenosis is the main defect.The function of the stent implantation is a sort of finit injury to the vascular stenosis,while the process of repairment is a common and basic vascular reaction, including the reconstruction and remodeling of local vascular and the migration, proliferation and apoptosis of vascular smooth muscle cell etc. which mediated by located inflammatory reaction,various of cytokines and growth factors. How to prevent the coronary restenosis after PCI is always a hot topic to global cardiovascular physicians.This article mainly studied the establishment of light initiated chemiluminescence assay (LiCA) for the quantitatively detection of human plasma MPO and the level changes of MPO, cTnI, hs-CRP, IL-6, WBC before and after stent implantation.Object: A light initiated chemiluminescence assay (LiCA) for the quantitative detection of human plasma MPO level was established to assist the diagnose of cardiovascular diseases.The level changes of MPO,cTnI, hs-CRP, IL-6 and WBC before and after coronary angiograms and PCI were separately observed.Further more,we have studied the changes of the related inflamatory factors in the patients'blood after PCI and their clinical value have also been explored.Methods: From twelve MPO McAbs, the optimum double-antibody combination (one coated on chemiluminescent particles and the other conjugated with biotin) were picked out . Then we explored the optimal concentration and conditions, established the standared curve.Subsequently, the repeat, sensetivity and recovery rate of the assay were evaluated, which were compared with the classic CardioMPO ELISA. Light initiated chemiluminescence assay(LiCA) is a quantitative method with the features of fast,avoiding washing,high sensitivity, easy operate and convinient by using homogeneous immune LED technology. The clinical data before and after coronary angiography and before and after cardiac stent implantation were separately compared,including gender,age and the level of MPO, cTnI, hs-CRP, IL-6, WBC, creatinine, uric acid, glucose, total cholesterol , triglyceride, low density lipoprotein cholesterol, high density lipoprotein cholesterol, homocysteine (Plasma MPO is tested by enzyme-linked immunosorbent assay CardioMPO and other indicators are received by the Second Military Medical University Department of Laboratory Diagnosis, Changhai Hospital). Statistical analysis was performed by using the software SPSS16.0 and EXCEL.Result: Two McAbs were selected successfully. Thus the MPO-LiCA system were constructed by connecting the sensitizer particles with strep tavidin. The results showed that the analytical sensitivity was 1.76 ng/ml. The intra-and inter-assay CV of MPO-LiCA were 2.73% and 3.76% respectively, and the mean recovery rate was 102.5%. No high-dose hook effect was observed at MPO concentrations up to 800 ng/ml.The corelation degree between the and CardioMPOTM Enzyme Immunoassay was highly related (r=0.961). The level of MPO in patients who suffered from Coronary angiography is not statistically significant(P=0.275),while the level of hs-CRP and IL-6 are separately statistically significant(P<0.001,P=0.013). The level of MPO and hs-CRP and IL-6 in those understaking stent implantation is statistically remarcable(p<0.001, P<0.001,P=0.018). Conclusion: we have successfully established a quantitative light initiated chemiluminescence assay which can be used to test plasma MPO. MPO-LiCA and the classic CardioMPO ELISA are basicly identical in quantity.Compard with the classic CardioMPOTM Enzyme Immunoassay, MPO-LiCA is more sensitivity,specifity,stability,practical ,wider scope and shows a great application prospect.The level of MPO do not change before and after Coronary angiography. But,the level of MPO is higher after stent implantation than before stent implantation.It demonstrates that the level of MPO is improving after stent implantation because of the injury coronary atery vessel , the gather of PMN and local inflammatory reaction,and it is independent of hs-CRP and IL-6.