A novel multiplex polymerase chain reaction assayfor profile analyses of gene expression in peripheralblood
Scritto da BMC Cardiovascular Disorders   
CardiologiaBackground:Studies have demonstrated that inflammation has a key role in the pathogenesis ofatherosclerosis due to the abnormal gene expressions of multiple cytokines. We establishedan accurate and precise method to observe gene expression in whole blood that might providespecific diagnostic information for coronary artery disease (CAD) and other related diseases.Methods:The fifteen selected CAD-related genes (IL1B, IL6, IL8, IFNG, MCP-1, VWF, MTHFR,SELL, TNFalpha, ubiquitin, MCSF, ICAM1, ID2, HMOX1 and LDLR) and twohousekeeping genes (ACTB and GK) as internal references have been measuredsimultaneously with a newly developed multiplex polymerase chain reaction (multi-PCR)MethodMoreover, the precision was evaluated, and a procedure for distinguishing patientsfrom the normal population has been developed based upon analyses of peripheral blood. Atotal of 148 subjects were divided into group A (control group without plaques), group B(calcified plaques) and group C (non-calcified plaques, and combination group) accordingdual-source CT criteria. Gene expression in blood was analyzed by multi-PCR, and levels ofglucose and lipids measured in 50 subjects to explore the relationship among them.Results:The precision results of the multi-PCR system revealed within-run and between-run CVvalues of 3.695-12.537% and 4.405-13.405%, respectively. The profiles of cytokine geneexpression in peripheral blood were set: a positive correlation between glucose and MCSF,HMOX1 or TNFalpha were found. We also found that triglyceride levels were negativelycorrelated with SELL gene expression in 50 subjects. Compared with controls, geneexpression levels of IL1B, IL6, IL8 and MCP-1 increased significantly in group C.Conclusions:A new multiple gene expression analysis system has been developed. The primary datasuggested that gene expression was related to CAD. This system might be used for riskassessment of CVDs and other related diseases.
Fonte: BMC Cardiovascular Disorders
 

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