title: Evaluation of High Resolution Melting for MTHFR C677T Genotyping in Congenital Heart Disease.
abstract: BACKGROUND
  High resolution melting (HRM) is a simple, flexible and low-cost mutation screening technique. The methylenetetrahydrofolate reductase (MTHFR) gene encoding a critical enzyme, potentially affects susceptibility to some congenital defects like congenital heart disease (CHD). We evaluate the performance of HRM for genotyping of the MTHFR gene C677T locus in CHD cases and healthy controls of Chinese Han population.
  
  
  METHODS
  A total of 315 blood samples from 147 CHD patients (male72, female 75) and 168 healthy controls (male 92, female 76) were enrolled in the study. HRM was utilized to genotype MTHFR C677T locus of all the samples. The results were compared to that of PCR-RFLP and Sanger sequencing. The association of the MTHFR C677T genotypes and the risk of CHD was analyzed using odds ratio with their 95% confidence interval (CIs) from unconditional logistic regression.
  
  
  RESULTS
  All the samples were successfully genotyped by HRM within 1 hour and 30 minutes while at least 6 hours were needed for PCR-RFLP and sequencing. The genotypes of MTHFR C677T CC, CT, and TT were 9.52%, 49.66%, and 40.82% in CHD group but 29.17%, 50% and 20.83% in control group, which were identical using both methods of HRM and PCR-RFLP, demonstrating the sensitivity and specificity of HRM were all 100%.
  
  
  CONCLUSIONS
  MTHFR C677T is a potential risk factor for CHD in our local residents of Shandong province in China. HRM is a fast, sensitive, specific and reliable method for clinical application of genotyping.
authors.affiliation: Research Institute of Pediatrics, Qilu Children's Hospital of Shandong University, Ji'nan, 250022, China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Research Institute of Pediatrics, Qilu Children's Hospital of Shandong University, Ji'nan, 250022, China.
authors.forename: Haiyan
authors.initials: H
authors.lastname: Zhang
authors.affiliation: Department of Medicine, Shandong Medical College, Ji'nan, 250002, China.
authors.forename: Shuying
authors.initials: S
authors.lastname: Yue
authors.affiliation: Cardiovascular Department, Qilu Children's Hospital of Shandong University, Ji'nan, 250022, China.
authors.forename: Kun
authors.initials: K
authors.lastname: Zhang
authors.affiliation: Cardiovascular Department, Qilu Children's Hospital of Shandong University, Ji'nan, 250022, China.
authors.forename: Hui
authors.initials: H
authors.lastname: Wang
authors.affiliation: Research Institute of Pediatrics, Qilu Children's Hospital of Shandong University, Ji'nan, 250022, China.
authors.forename: Rui
authors.initials: R
authors.lastname: Dong
authors.affiliation: Research Institute of Pediatrics, Qilu Children's Hospital of Shandong University, Ji'nan, 250022, China.
authors.forename: Xiaomeng
authors.initials: X
authors.lastname: Yang
authors.affiliation: Research Institute of Pediatrics, Qilu Children's Hospital of Shandong University, Ji'nan, 250022, China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Research Institute of Pediatrics, Qilu Children's Hospital of Shandong University, Ji'nan, 250022, China.
authors.forename: Yanhui
authors.initials: Y
authors.lastname: Ma
mesh_terms: D002675:Child, Preschool; D005260:Female; D005838:Genotype; D006330:Heart Defects, Congenital / Q000201:enzymology / Q000453:epidemiology / Q000235:genetics*; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D008297:Male; D042965:Methylenetetrahydrofolate Reductase (NADPH2) / Q000235:genetics*; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length
chemical_list: C573423:MTHFR protein, human; D042965:Methylenetetrahydrofolate Reductase (NADPH2)