Duplex Allele-Specific PCR Assay Simultaneously Detecting Genotype of Osteoarthritis Susceptibility Loci Within GDF5 and COL11A1 Genes

Author(s): Svetlana B. Panina, Igor V. Kornienko, Igor V. Krolevets, Mikhail A. Zabrodin, Natalia P. Milyutina

Introduction: Genome-wide association studies and meta-analyses have identified rs143383 GDF5 (growth and differentiation factor 5) and rs2615977 COL11A1 (collagen type XI alpha 1) as polymorphic loci among the most robust associations with osteoarthritis (OA). However, currently, there is a lack of available rapid, technically feasible, and costeffective methods to detect OA susceptibility genotype, not linked with the specific descent of the patients.

Methods: Genomic DNA was extracted from the blood of OA patients and healthy individual using standard methods. Primers specific at the 3'-end for the polymorphic sites were designed. Direct sequencing was used to determine the genotype of the study subjects.

Results and conclusion: Here we describe one-step allele-specific PCR assay simultaneously detecting genotype of rs143383 GDF5 and rs2615977 COL11A1. Duplex PCR enabled to obtain specific bands corresponding to the amplified product of interest. The genotypes called based on PCR results were consistent with the results of direct sequencing.

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