Science. Reliable identification of the type III collagen gene polymorphism rsl800255 with the use of high resolution melting analysis.

Background: We have recently identified a homozygous nucleotide substitution in exon 30 of type III collagen (COL3A1 2209G>A, rs1800255) as a risk factor for the development of pelvic organ prolapse. This polymorphism is also associated with mitral valve prolapse. Patients could benefit from preventive actions when identified prior to clinical evidence of the disease. High resolution melting (HRM) analysis is a low-cost, high-throughput technique to identify DNA polymorphisms. The purpose of this study was to investigate whether HRM is an accurate method to detect the above mentioned polymorphism in DNA from blood samples Methods: A total of 36 randomly selected DNA samples from pelvic organ prolapse patients were analyzed in duplicate using polymerase chain reaction (PCR)-based restriction fragment length polymorphism (PCR-RFLP) and sequence analysis. Using the latter as the gold standard, all samples were subjected to HRM analysis with the use of specific primers and an unlabelled probe. Results: Compared to sequence analysis, PCR-RFLP resulted in 2 incorrect diagnoses. High resolution melting analysis demonstrated unique melting profiles for wild-type, as well as heterozygous and homozygous polymorphisms, with 100% accuracy. Conclusion: High resolution melting is a fast, cost-effective, and sensitive screening method to detect the rs1800255 polymorphism. [ABSTRACT FROM AUTHOR]