Comparative analyses of DNA extraction methods for whole blood quantification of HCMV DNAemia in patients with hematological diseases: false negative cases in manual method

Gabriella Bianchino; Vitina Grieco; Giuseppe Pietrantuono; Sabino Russi; Luigi Del Vecchio; Geppino Falco; Tiziana Notarangelo

doi:10.1016/j.heliyon.2023.e18740

Comparative analyses of DNA extraction methods for whole blood quantification of HCMV DNAemia in patients with hematological diseases: false negative cases in manual method

Heliyon. 2023 Jul 27;9(8):e18740. doi: 10.1016/j.heliyon.2023.e18740. eCollection 2023 Aug.

Authors

Gabriella Bianchino¹, Vitina Grieco¹, Giuseppe Pietrantuono¹, Sabino Russi¹, Luigi Del Vecchio^{2

3}, Geppino Falco^{4

5}, Tiziana Notarangelo¹

Affiliations

¹ IRCCS CROB, Centro di Riferimento Oncologico della Basilicata, 85028, Rionero in Vulture (PZ), Italy.
² Biotecnologie Avanzate, CEINGE, Naples, Italy.
³ Department of Molecular Medicine and Medical Biotechnologies, "Federico II" University of Naples, Italy.
⁴ Department of Biology, University of Naples Federico II, 80126, Naples, Italy.
⁵ Biogem, Istituto di Biologia e Genetica Molecolare, Via Camporeale, 83031, Ariano Irpino (AV), Italy.

Abstract

Human cytomegalovirus (HCMV) DNA quantitation in whole blood (WB) by real-time or quantitative polymerase chain reaction (qPCR) is a highly sensitive and reproducible diagnostic procedure for monitoring HCMV DNAemia (DNAemia is the detection of DNA in samples of plasma, whole blood, isolated peripheral blood leukocytes or in buffy-coat specimens) in patients. We provided a comparative analysis of HCMV DNA extraction performance by two different techniques, one performed by an automated extractor and the other by a manual method. We observed that the automated extraction method allowed HCMV DNA detection in the presence of weak viremia while no differences are observed when the viral load is greater. Therefore, automated DNA extraction is a suitable and recommended protocol not only for early detection of HCMV infection but also for more accurate monitoring of HCMV DNAemia during post-therapy follow-up.

Keywords: DNA extraction; HCMV; Hematological diseases.