Quantification of human T-cell lymphotropic virus type 1 proviral load by quantitative competitive polymerase chain reaction

The polymerase chain reaction (PCR) has been established as a highly sensit ive technique for detection of viral DNA or RNA. However, due to inherent l imitations of PCR the amount of amplified product often does not correlate with the initial amount of template DNA. This is particularly true for PCR detection of viral infections that are characterized by low in vivo viral c opy numbers in certain stages of the infection, such as human T-cell lympho tropic virus type 1 (HTLV-1) and simian T-cell lymphotropic virus type 1 (S TLV-1). Therefore, we developed a quantitative competitive polymerase chain reaction (qcPCR) for detection of HTLV-1 and STLV-1 proviral DNA. The assa y was optimized using an infectious HTLV-1 clone, AGH, HTLV-1 infected cell lines, MT-2.6 and HUT-102 and STLV-1 infected lines Kia and Matsu. Applica bility of this system was demonstrated by determining HTLV-1 proviral load in peripheral blood mononuclear cells (PBMC) of human subjects with HTLV-1 associated diseases and an asymptomatic carrier as well as rabbits infected experimentally. This qcPCR method, the first designed specifically for HTL V-1 and STLV-1, will provide an important tool for pathogenesis studies of HTLV-1 and for evaluating the efficacy of antiviral drugs and vaccines agai nst the viral infection using animal models. (C) 1998 Elsevier Science Irel and Ltd. All rights reserved.