Complement C4 Gene Copy Number Variation Genotyping by High Resolution Melting PCR.

Area of Science:

• Genetics
• Molecular Biology

Background:

• Complement C4 gene copy number variation (CNV) is linked to susceptibility in autoimmune diseases like lupus, schizophrenia, and rheumatoid arthritis.
• Accurate quantification of C4 CNVs is essential for understanding disease pathogenesis and genetic risk.

Purpose of the Study:

• To develop and validate a novel assay for quantifying complement C4 gene copy number variations.
• To simplify the diagnostic process for C4 CNV.

Main Methods:

• The assay utilizes gene ratio analysis copy enumeration (GRACE) PCR combined with high-resolution melting (HRM) PCR.
• Optimization involved samples with known genotypes, followed by validation using 72 DNA samples from healthy donors.

Main Results:

• Standard curves were generated by plotting C4/RP1 ratio values against copy number variation (CNV) for each gene.
• The observed range of C4 copy numbers in control individuals aligns with previous findings in European populations.

Conclusions:

• The developed method offers a significant simplification for diagnosing complement C4 copy number variations.
• This assay facilitates more accessible and efficient C4 CNV analysis.