An Improved High Resolution Melting Curve For MTHFR C677T Detection

Objective: To establish an improved protocol based on high resolution melting curve(HRM)and apply it to methylenetetrahydrofolate reductase(MTHFR)gene C677 T polymorphism(rs1801133)detection.Methods: A pair of universal primers with the M13 linker was designed for the MTHFR C677 T.To optimized its analysis performance,HRM reaction system was improved.The M13 linker was used as a Sanger sequencing primer to directly sequence the HRM products in order to verify HRM results.The improved protocol was applied to detect the MTHFR C677 T single nucleotide polymorphism of 1030 peripheral blood samples which were collected from our hospital from 2016.06 to 2017.10.Results: The improved HRM was successfully established and applied to MTHFR C677 T polymorphism detection.Among 1030 deoxynucleic acid(DNA)samples,CC genotype accounted for 39.13%,CT genotype accounted for 48.16% and TT genotype accounted for 12.71%.Conclusion: A genotyping method for detecting MTHFR C677 T was established by improving HRM.The method proved a simple,economical,rapid,accurate and high-throughput tool for large-scaleclinical genetic detection.