Usage Of Suspension Chip Integrated Detection Technique In Prewarning Of Acute Coronary Syndrome

Acute coronary syndrome (ACS) attacks because of atherosclerotic plaque instability, plaque ulceration, rupture, leading to the formation of blood clots caused by a severe acute ischemic heart disease, including unstable angina pectoris (unstable angina pectoris, UAP), non-ST-segment elevation myocardial infarction (non-ST elevation infarction, NSTEMI) and ST-segment elevation myocardial infarction (ST elevation infarction, STEMI) and sudden cardiac death, compared with stable angina pectoris, patients of ACS whose clinical syndrome is characterized by fast change of condition, often lack the protection measures, high mortality, difficult to predict patients'prognosis and so on. So its early diagnosis and timely treat has a significant meaning. Currently the "gold standard" of diagnosis of CHD is coronary angiography, which can be more understandable of the extent and scope of coronary artery lesion, but as a kind of invasive examination, it has a certain degree of risk and cost more. it can not be used as routine diagnostic examination, Therefore it is necessary to find a simple and quick diagnostic method that make us have a clear diagnosis of ACS.ACS is a multi-links disease caused by various pathogenic factors, whose recognized risk factor include smoking, obesity, and hypertention, but these factors can not fully explain the causes and severity of coronary heart disease. In recent years, some new risk factor for coronary heart disease is increasingly concerned by researchers. The current evidence suggests that inflammation is one of the related factor caused the formation and development of atherosclerotic plaque, and studies confirmed the persistent inflammation reaction also exist in the atherosclerotic plaques. Acute inflammatory reaction is not only an important pathological mechanism of atherosclerotic plaque instability, but also correlate to the severity and prognosis of ACS. Therefore, inflammatory factors may predict the potential risk of coronary disease. In present days vulnerable plaque has become the focus of ACS reasearch, which mainly refers to the plaque prone to instability and thrombosis, including plaque rupture, plaque erosion and calcified nodules.Suspension microarray technology is a new technology of protein chip, the method was beneficial to the processing of photoelectric conversion and digital signal technology, which use suspended microsphere in the reaction solution to overcome the problems of membrane chip-chip detection such as influnce on surface tension, spatial effect on the reaction kinetics, contamination and so on. AS it rely on the laser detection, identification code of the micorsphere and the fluorescence signal biological responses, make it has many adavantages such as high-throughput, simple, reproducible, high sensitivity and wide dynamic range and so on. It has been widely used in tissue typing, genetic disease diagnosis, cancer, genetic testing, the detection of SNP and mRNA/cDNA expression detection.ObjectiveTo investigate the feasibility of integrated detection for acute coronary syndrome (ACS)-associated protein factors [high sensitive c-reactive protein(hs-CRP), lipoprotein(a)[Lp(a)], adiponectin(APN)] by Microsphere Chip, and try to analysis the relationship between the three factors [hs-CRP, Lp(a) and APN]and ACS.Subjects and Method1. Subjects96 patients (64 patients with acute myocardial infarction, unstable angina 32 patients) were selected diagnosed as acute coronary syndrome in department of cardiology, zhujiang hospital from March 2008 to December 2008,including 60 males and 36 females. Their mean age is 63.11±10.84 years. There are 30 patients of stable angina which include 18 males and 12 females, mean age (59.86±10.54) years and n 40 healthy subjects includ 17 males and 23 female, mean age (62.02±10.11) years old was slected as control.All the subjects were drawn 5mL peripheral blood. The serum samples were segregated and preserved in-80℃refrigerator.2. Method2.1 Give all the ACS patients without contraindications standard secondary prevention of coronary heart disease. All the ACS patients were receiving emergency or elective coronary angiography and appropriate revascularization. Give antiplatelet therapy with aspirin, clopidogrel heparin or plus tirofiban after the intervention surgery.2.2 General information must be registered with:age, sex, smoking and other risk factors, disease risk stratification and general laboratory findings.2.3 All the ACS patients were followed up for cardiovascular events, such as arrhythmia, heart failure, stroke, and recurrence of angina/Ml, cardiac death.2.4 To establish an integrated quantitative detection method by microsphere chip for hs-CRP, Lp(a) and APN.2.5 These three factors from the serum samples were detected by microsphere chip and enzyme-linked immunosorbent assay (ELISA) meanwhile. Then compared the result of two methods.2.6 Compared the serum concentration of three protein factors in different groups and approach the relationship between the three factors and ACS.2.7 To study the value of integrated detection by microsphere chip in the anticipation and diagnosis of ACS, Logistic regression analysis was done for the anticipation of prognosis. 3. Statistical analysisSPSS 13.0 statistical soft was used. The measurement data was shown as mean±SD (x±S). The compared of multi-groups was tested by one-way ANOVA after homogeneity of variance test, and multiple comparison in different groups was tested by LSD. If equal variances not assumed or the data obey skewness distribution, the compared of multi-groups was tested by Krushal-Wallis H test, and multiple comparison in different groups was tested by Neminyi Test. The count data was shown as frequency. The compared of multi-groups was tested byχ2 test. Spearman correlation analysis was used in skewness data bivariate correlation analysis. Binary Logistic Regression Analysis was used in regression analysis. Statistical significance is defined as P<0.05 on two-tailed testing.Results1. There was no significant difference in the general information among these groups.2. The detection results by microsphere chip integrated detection method correlate significantly to that of ELISA (hs-CRP:rs=0.934, P< 0.001; Lp(a): rs=0.909, P< 0.001; APN:r,=0.843,P< 0.001). 3. There was significant difference in the levels of hs-CRP, Lp(a) and APN among these groups(P<0.01). The level of hs-CRP in ACS group was significant higher than that in Control group and SAP group, and AMI sub-group of poor prognosis have a higher CRP and Lp (a) concentrations than in UAP sub-group of good prognosis, APN were significantly lower than patients with UAP sub-group of good prognosis (P all<0.001).4. Compared with NC group, hs-CRP, Lp (a) in SAP was higher than NC (P all<0.05), there was no significant difference between two groups.(P> 0.05)5. AMI and UAP group with short term and long-term poor prognosis have a higher level of CRP and Lp (a) concentrations,and a lower level of APN than those with good porgnosis, APN (P all<0.05).6. Regression analysis of shows:regression equation was y=-10.307 +1.556 x1+0.234 x2+0.003 x3-0.292x4, equation sentenced rate are 96.9%. hs-CRP, partial regression coefficient of 0.234, standardized regression coefficient 0.09, P <0.01, OR value of 1.264; LP (a), partial regression coefficient 0.003, standard regression coefficient 0.001, P<0.05, OR value of 1.003; APN, partial regression coefficient,-0.292, standard regression coefficient 0.122, P<0.05, OR value of 0.746.Conclusion1.There was no significant difference of age, sex, weight, etc. between each groups.2, Established the quantitative detection method of coronary heart disease suspended chip, which include simultaneous detection of hs-CRP, Lp (a) and APN.3, successfully suspension array technology for protein factors associated with acute coronary syndrome testing, and proved suspended chip multi-factor quantitative detection method.4, using suspension chip integrated detection of acute coronary syndrome related protein factors and the results are beneficial to a comprehensive analysis of early diagnosis of acute coronary syndrome in order to further address detection technology to achieve accurate, rapid, simple, multi-function, multi-target, high flux, the requirements of standardized basis.5, hs-CRP, Lp (a) is an important marker of acute inflammation, acute coronary syndrome were independent risk factors, participants in the inflammatory activity within atherosclerotic plaque, and the poor prognosis group in both ACS were significantly higher than the prognosis is better, therefore risk of coronary heart disease occurs as an important prognostic predictor. APN levels were significantly lower and stable angina, and poor prognosis in the ACS group were significantly lower than those with good prognosis, suggesting that inflammation has a protective effect against coronary artery.6 Multivariate logistic regression analysis showed that, hs-CRP, Lp (a) is a recent acute coronary syndrome risk factors of poor prognosis, prompt intervention with plaque progression after treatment, new lesions, but APN was protective factors.