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Amplifying and Quantifying HIV-1 RNA in HIV Infected Individuals with Viral Loads Below the Limit of Detection by Standard Clinical Assays
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Whole Blood Volume-Based Absolute Quantification of HTLV-1 Proviral Load: A Comparative Method Evaluation Study.

Gabriel O Franco1,2,3, Andreas Stocker1,2,3, Eduardo M Netto1,2

  • 1Laboratório de Pesquisa em Infectologia (LAPI), Complexo Hospitalar Universitário Professor Edgard Santos, Salvador 40110-060, Brazil.

Viruses
|May 27, 2026
PubMed
Summary

Quantifying human T-cell lymphotropic virus type 1 (HTLV-1) proviral load using whole blood volume is a simpler, effective alternative. This method aids in monitoring adult T-cell leukemia/lymphoma (ATL) and HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP).

Keywords:
HTLV-1absolute quantificationmethodological standardizationproviral loadqPCR

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Area of Science:

  • Virology
  • Immunology
  • Molecular Diagnostics

Background:

  • Proviral load of human T-cell lymphotropic virus type 1 (HTLV-1) is a key biomarker for monitoring adult T-cell leukemia/lymphoma (ATL) and HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP).
  • Current quantification methods lack standardization, hindering broader clinical application and risk stratification.

Purpose of the Study:

  • To evaluate a novel absolute quantification approach for HTLV-1 proviral load based on whole blood volume.
  • To compare the performance of volumetric quantification with established protocols, including cell-normalized methods.

Main Methods:

  • Analysis of 66 HTLV-1-infected individuals using six quantitative PCR (qPCR) methodologies.
  • Comparison of volumetric quantification (copies/µL) with Tamegão-Lopes method and cell-normalized counts (per 1000 cells) in whole blood, buffy coat, PBMCs, and CD4+ T cells.
  • Statistical assessment of association and agreement using correlation, Bland-Altman analysis, CCCs, and Deming regression.

Main Results:

  • Absolute quantification based on whole blood volume demonstrated strong correlation (r=0.93) and high agreement (CCC=0.866) with the Tamegão-Lopes method.
  • Volumetric quantification also showed strong correlation (r=0.84) and high agreement (CCC=0.811) with CD4+-based quantification.
  • Leukocyte-normalized methods exhibited greater discrepancies, likely due to dilution effects from uninfected cells.

Conclusions:

  • Quantification of HTLV-1 proviral load based on total blood volume offers a simplified and operationally feasible alternative.
  • This volumetric approach provides a reliable method for assessing HTLV-1 proviral load, supporting clinical monitoring and risk stratification.
  • Standardization of HTLV-1 proviral load quantification can be improved through volumetric methods, enhancing their utility in managing HTLV-1-associated diseases.