[Rapid screening for MTHFR gene 677C>T polymorphism in Down syndrome using high resolution melting curve and...

Area of Science:

• Molecular genetics
• Genetic polymorphism analysis
• Biotechnology

Background:

• The methylenetetrahydrofolate reductase (MTHFR) gene 677C>T polymorphism is clinically significant.
• Accurate and rapid detection methods are crucial for genetic studies.

Purpose of the Study:

• To develop and compare high-resolution melting curve (HRM) and pyrosequencing methods for MTHFR gene 677C>T polymorphism detection.
• To evaluate the accuracy and efficiency of these methods for genotypic and allelic frequency determination.

Main Methods:

• Comparison of regular HRM, internal control HRM, and artificial heterozygosity HRM.
• Pyrosequencing analysis of allele frequencies in mixed samples.
• Validation using peripheral blood samples from Down syndrome patients and normal controls.

Main Results:

• Internal control and artificial heterozygosity HRM effectively distinguished CC, CT, and TT genotypes.
• Pyrosequencing accuracy for allele frequency improved with sample size, achieving <4% difference with >30 samples.
• TT genotype was more prevalent in Down syndrome patients (25.2%) compared to controls (14.3%).

Conclusions:

• Internal control HRM is suitable for genotypic frequency determination.
• Pyrosequencing is an ideal method for allelic frequency determination.
• These methods provide rapid and accurate tools for MTHFR gene polymorphism analysis.