ABSTRACT
Objectives@#The study aimed to validate a high resolution melting (HRM) – polymerase chain reaction (PCR) based method for analysis of ATP2B1 rs2681472, a candidate genetic marker for hypertension. Specifically, the study aimed to establish method accuracy, inter- and intra-day precision, instrument detection limits (IDL) and linearity. @*Methodology@#DNA samples from whole blood of selected respondents of the 2008 National Nutrition Survey (NNS) were analyzed for ATP2B1 rs2681472, following the HRM-PCR method. Analytical validation parameters such as method accuracy, inter- and intra-day precision, IDL and assay linearity were evaluated. @*Results@#The data obtained using HRM conformed to data obtained from the current gold standard, which is direct DNA sequence analysis. Precision of the method is shown in the low intra-day and inter-day coefficients of variation of 0.11% and 0.14% respectively. The IDL was found at 1x10-2 ng DNA. Correlation coefficient using the method has met the acceptance criteria for linearity that is equal to 0.98. @*Conclusion@#HRM-PCR was successfully applied for analysis of ATP2B1 rs2681472. HRM is a highly commendable tool that can identify existence of polymorphisms in a genome which may be related to or which may cause hypertension. The method was found to be accurate and precise.
Subject(s)
HypertensionABSTRACT
Objective To investigate the association between polymorphism of ATP2B1 gene,its interaction with smoking and susceptibility of essential hypertension. Methods A case-control study was conducted to elucidate the role of ATP2B1 gene variants related to the risk of essential hypertension. Genomic DNA was extracted from peripheral blood leukocytes,using the QIAamp DNA Mini Kit(QIAGEN,Germany). Two SNPs,-rs17249754 and rs6253,were examined on 1 280 patients and 1 010 healthy controls,using a Snapshot method. Statistical analyses were performed with SPSS Windows software(version 19.0;SPSS,Chicago,IL). Results A significant difference was found in rs17249754 allele frequency between cases and controls(OR=1.223,95%CI:1.083-1.381,P=0.001). After adjustment for age,sex,BMI,smoking and drinking,the difference was still statistically significant(OR=1.212,95%CI:1.070-1.373,P=0.003). In addition,data from genotype distribution analysis under different models showed that appeared significant associations between ATP2B1 gene polymorphism and essential hypertension(additive model OR=1.469,95%CI:1.121-1.925,P=0.005;dominant model OR=1.324,95%CI:1.029-1.704,P=0.029;recessive model OR=1.123,95%CI:1.031-1.223,P=0.008). In this study,the proportion of smokers in cases was significantly higher than that in controls(P=0.005),but no associations between rs17249754-smoking interaction and essential hypertension were found after the adjustment for gender,age,BMI and alcohol consumption (OR=1.024,95%CI:0.614-1.707). Conclusion Our research findings showed that the polymorphism of ATP2B1 gene rs17249754 was significantly associated with the incidence of essential hypertension in Han population of northeastern China. However,the interaction between rs17249754 and smoking did not seem to have contributed to the occurrence of the essential hypertension.
ABSTRACT
Objective To investigate the association between polymorphism of ATP2B1 gene,its interaction with smoking and susceptibility of essential hypertension. Methods A case-control study was conducted to elucidate the role of ATP2B1 gene variants related to the risk of essential hypertension. Genomic DNA was extracted from peripheral blood leukocytes,using the QIAamp DNA Mini Kit(QIAGEN,Germany). Two SNPs,-rs17249754 and rs6253,were examined on 1 280 patients and 1 010 healthy controls,using a Snapshot method. Statistical analyses were performed with SPSS Windows software(version 19.0;SPSS,Chicago,IL). Results A significant difference was found in rs17249754 allele frequency between cases and controls(OR=1.223,95%CI:1.083-1.381,P=0.001). After adjustment for age,sex,BMI,smoking and drinking,the difference was still statistically significant(OR=1.212,95%CI:1.070-1.373,P=0.003). In addition,data from genotype distribution analysis under different models showed that appeared significant associations between ATP2B1 gene polymorphism and essential hypertension(additive model OR=1.469,95%CI:1.121-1.925,P=0.005;dominant model OR=1.324,95%CI:1.029-1.704,P=0.029;recessive model OR=1.123,95%CI:1.031-1.223,P=0.008). In this study,the proportion of smokers in cases was significantly higher than that in controls(P=0.005),but no associations between rs17249754-smoking interaction and essential hypertension were found after the adjustment for gender,age,BMI and alcohol consumption (OR=1.024,95%CI:0.614-1.707). Conclusion Our research findings showed that the polymorphism of ATP2B1 gene rs17249754 was significantly associated with the incidence of essential hypertension in Han population of northeastern China. However,the interaction between rs17249754 and smoking did not seem to have contributed to the occurrence of the essential hypertension.