A quantitative real-time PCR assay has been practically used to measure the HTLV-1 proviral load in PBMCs, subsequently making a diagnosis of persistent HTLV-1 infection and HTLV-1-associated disorders, and monitoring cell burden, namely, the numbers of lymphocytes or ATL cells harboring the HTLV-1 genome within their genomic DNA. These assays have been described to be excellent for diagnostic sensitivity and specificity in many articles cited using a Medline search. Our PCR assay also has given us a reasonable outcome [i.e., all of the seronegative samples were undetectable, whereas the seropositive samples were all positive in 108 cases without ATL (healthy carriers) and in 59 cases with overt ATL]. The HTLV-1 proviral load was 301 ±339 copies (mean±SD) per 104 PBMCs in healthy carriers, and ranged from 1.72 x 102 to 1.53 x 104 copies in ATL in parallel with ATL cell counts. Of the 108 healthy carriers who had no clinical manifestations without a clonal band by the HTLV-1 SBH test, the proviral load, in combination with the percentage (%) of ATL-like flower cells defined by PB smear morphology, enabled the carriers to be subgrouped into three categories  as shown in Fig. 3. However, in 7 of 59 ATL cases characterized by the SBH test for HTLV-1 integration status, the proviral load and ATL cell counts by morphology were discrepant. Interestingly, all of these cases showing discrepancies had two or three bands by the HTLV-1 SBH analysis using the EcoRI restrictive enzyme which has no cleavage site within the proviral DNA sequence. The ratio of the proviral load to ATL cell number was closely equivalent to the band number obtained by the SBH analysis, indicating that ATL cells from these cases harbored multicopies within a single ATL cell.
These cross-sectional data from each real-time PCR assay system for HTLV-1 quantification have been generally reported to be concordantly stable and significant, but the true usefulness of the ''quantification'' of
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