The role of Single Nucleotide Variants (SNVs) / Single Nucleotide Polymorphisms (SNPs) in progression and cause of various chronic diseases, including Coronary Heart Diseases and other conditions is well documented. The present known techniques for the identification of genetic mutations, such as RFLP and qRT-PCR are very limited in their capacity that can identify not more than one mutation in a single run. Hence, a comprehensive genetic analysis of multiple mutations becomes labour-intensive and time-consuming.
On the other hand, highly informative and advanced methods such as DNA sequencing are not suitable for routine, small-scale diagnostic laboratories; since high sample throughput are required for NGS / Sanger sequencing methods to be economically viable, thus becomes very expensive for routine utility.
Hence, there is an imperative need for the development of a cost-effective method need for quick and effective genetic screening of the well-known/ well documented mutations for Cardiovascular Diseases. By utilizing a single, multiplex amplification reaction, the known genetic mutation markers can be detected within a few hours from blood drawing to final results. The number of mutations covered this assay can easily be extended to up to 48 mutations using the same approach to select additional hybridization probes.