Establishment of HTLV-1 gene quantification method using real-time PCR

Abstract

In organ transplantation, recipients are known to experience serious complications from infections after transplantation from viruses brought by organ donors. Therefore, it is important for donors to confirm the presence of infection in pre-transplantation testing for infectious diseases that are contraindicated for transplantation, such as human T-cell leukemia virus type 1 (HTLV-1). However,in living donor renal transplantation, there is no contraindication to organ donation from an HTLV-1-infected donor for recipients who were infected with HTLV-1 prior to transplantation. Therefore,authors believe that quantification of HTLV-1 is necessary to assess the risk of infection. In this study, we aimed to establish a quantitative real-time polymerase chain reaction (quantitative PCR) method that can be used as a definitive method for rapid recipients and donors diagnosis, assuming cases in which the decision was withheld by the line blotting assay (LIA) method. The standard curve required for quantitative PCR was calculated as HTLV-1 DNA copies/μL from threshold cycle (Ct) values using DNA fragments generated from HTLV-1 sequences. The quantitative PCR established in this study uses artificially created DNA fragments, which should enable stable quantification of HTLV-1.