Optimization of PCR DNA Sequencing Method for SNP Detection in Abacavir Sensitivity Gene - Abstract
Background: Abacavir is an antiretroviral indicated in combination with other antiretroviral agents for the treatment of HIV-1 infection. It is associated with potentially serious hypersensitivity reactions in 3–9% of Caucasian patients. Carriers of HLA-B*5701 are at high risk for developing abacavir hypersensitivity reaction (AHR). In Caucasians, HCP5 rs 2395029(G) is in high but not complete linkage disequilibrium (LD) with HLA-B*5701.
Aim: To development of fast method to screening of HLA-B*5701 allele, by optimization of PCR DNA Sequencing method for SNP detection in Abacavir Sensitivity gene.
Methods: DNA extraction by whole blood Sample using the QIAamp® DNA Blood mini kit (Qiagen, Hilden, Germany). HLA-B*5701 allele Specific amplification Primer is design in Primer3 ( online tool ) using db SNP ID-rs2395029 as Reference Sequences, these Primer is used PCR and PCR DNA Sequencing . SNP detected by Chromas Software using db SNP ID-rs2395029 as Reference Sequences.
Results: We propose a simple approach to detection of SNP of HLA-B*5701 allele associated abacavir hypersensitivity based on optimization of PCR DNA Sequencing Method of HLA-B*5701 allele to determine the presence of hypersensitivity risk.
Conclusion: HLA-B*5701 genotyping before initiation of therapy remain probative of .the Introduction of highly active antiretroviral therapy has Transformed the nature of HIV Infection from a severe and ultimately fatal disease to that of a manageable chronic condition. HIV drugs are highly efficacious, but there use comes at the cost of a range of drug –related adverse events, including sever hypersensitivity reaction (HSRs) that have been most notably associated with abacavir and other NRTIs drug therapy.