Validation of real-time quantitative RT-PCR assay.

(A) To determine interassay CV% for this real-time RT-PCR assay, cDNA from the HTLV-1–infected MT-2 cell line was serially diluted with cDNA from the uninfected Jurkat cell line and HTLV-1 tax mRNA load was measured 5 times per dilution. (B) The intra-assay CV% was determined by measuring mRNA load from the PBMCs of 3 HAM/TSP patients in 5 separate experiments. The CV% represents (SD of HTLV-Itax mRNA load/average of HTLV-I tax mRNA load) × 100.