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Quantitation of human herpesvirus-6A, -6B and -7 DNAs in whole blood, mononuclear and polymorphonuclear cell fractions from healthy blood donors
Authors:Bastien Gé  raudie,Mé  linda Charrier,Pascale Bonnafous,Delphine Heurté  ,Marion Desmonet,Marc-Antoine Bartoletti,Cé  cile PenasseHenri Agut,Agnè  s Gautheret-Dejean
Affiliation:a UPMC Univ Paris 06, ER1 DETIV, 83 Boulevard de l’Hôpital, Paris F-75013, France
b Service of Virology, Pitié-Salpêtrière Hospital, AP-HP, 83 Boulevard de l’Hôpital, Paris F-75013, France
c French Blood Bank (EFS), site Pitié-Salpêtrière, Pitié-Salpêtrière Hospital, AP-HP, 83 Boulevard de l’Hôpital, Paris F-75013, France
d Laboratory of Microbiology, UPRES EA 4065, Paris Descartes University, Faculty of Pharmacy, 4 Avenue de l’Observatoire, Paris F-75006, France
Abstract:

Background

Diagnosis of human herpesvirus-6A (HHV-6A), -6B (HHV-6B) or -7 (HHV-7) infections is often based on the measure of viral load in blood.

Objectives

The aim of this study was to define usual values of HHV-6A, HHV-6B and HHV-7 loads in blood fractions (whole blood [WB], mononuclear cells [PBMCs], polymorphonuclear leukocytes [PMNLs]) of blood donors.

Study design

HHV-6A, HHV-6B and -7 DNAs were quantitated using real-time PCR assays in WB, PBMCs and PMNLs separated on Ficoll or dextran gradients, respectively, for 200 blood donors. Viral loads were expressed as the number of viral genomic copies per million cells (Cop/M) for all fractions, and also per milliliter for WB.

Results

HHV-6B DNA was rarely detected in WB (8%), PBMCs (16.5%), and PMNLs (10.5%), HHV-6A was never detected, whereas HHV-7 DNA was often present in WB (51.5%), PBMCs (62%) and PMNLs (51.5%). Median loads were low with 81 Cop/M in WB, 62 Cop/M in PBMCs and 34.5 Cop/M in PMNLs for HHV-6B, and 129 Cop/M in WB, 225 Cop/M in PBMCs and 62 Cop/M in PMNLs for HHV-7. Viral load expression per million cells and per mL were equivalent. One subject had chromosomally integrated HHV-6 with high viral loads ranging from 2.23 × 106 to 3.21 × 106 Cop/M in all compartments and plasma.

Conclusions

These results allow to propose viral load in WB as a sensitive and suitable marker, with values for healthy subjects at approximately 100 Cop/M for both viruses. The prevalence of chromosomally integrated HHV-6 was 0.5%.
Keywords:Human herpesvirus-6   Human herpesvirus-7   Viral load   Whole blood   Peripheral mononuclear cells   Polymorphonuclear leukocytes   Blood donors
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