A novel multiplex polymerase chain reaction assay for profile analyses of gene expression in peripheral blood

被引:5
作者
Jia, Xingwang [1 ]
Ju, Haiyue [2 ]
Yang, Li [2 ]
Tian, Yaping [1 ]
机构
[1] Chinese Peoples Liberat Army Gen Hosp, State Key Lab Kidney Dis, Dept Clin Biochem, Beijing 100853, Peoples R China
[2] Chinese Peoples Liberat Army Gen Hosp, Dept Radiol, Beijing 100853, Peoples R China
基金
国家高技术研究发展计划(863计划); 中国国家自然科学基金;
关键词
Coronary artery disease; Gene expression profiling; Multiplex polymerase chain reaction; GeXP; MESSENGER-RNA EXPRESSION; MONICA PROJECT; CORONARY; MORTALITY; DISEASE; ATHEROSCLEROSIS; INFLAMMATION; TOMOGRAPHY; PATTERNS; REVEALS;
D O I
10.1186/1471-2261-12-51
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
Background: Studies have demonstrated that inflammation has a key role in the pathogenesis of atherosclerosis due to the abnormal gene expressions of multiple cytokines. We established an accurate and precise method to observe gene expression in whole blood that might provide specific 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 two housekeeping genes (ACTB and GK) as internal references have been measured simultaneously with a newly developed multiplex polymerase chain reaction (multi-PCR) method. Moreover, the precision was evaluated, and a procedure for distinguishing patients from the normal population has been developed based upon analyses of peripheral blood. A total 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) according dual-source CT criteria. Gene expression in blood was analyzed by multi-PCR, and levels of glucose 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 CV values of 3.695-12.537% and 4.405-13.405%, respectively. The profiles of cytokine gene expression in peripheral blood were set: a positive correlation between glucose and MCSF, HMOX1 or TNFalpha were found. We also found that triglyceride levels were negatively correlated with SELL gene expression in 50 subjects. Compared with controls, gene expression 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 data suggested that gene expression was related to CAD. This system might be used for risk assessment of CVDs and other related diseases.
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页数:9
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