Enhancing the sensitivity of HIV DNA detection: a magnetic bead-based method

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Enhancing the sensitivity of HIV DNA detection: a magnetic bead-based method

Journal Title: Chinese Journal of Blood Transfusion - Year 2023, Vol 36, Issue 12

Abstract

Objective To establish a magnetic bead enrichment strategy for the detection of human immunodeficiency virus deoxyribonucleic acid (HIV DNA) in peripheral blood, and to verify the improvement of the sensitivity of this method for the detection of HIV DNA in HIV infected patients after early antiretrovital treatment (ART). Methods Peripheral whole blood was collected at 4 timepoints in one ART HIV window period (WP) patient. Peripheral blood mononuclear cells (PBMCs) were isolated on a Ficoll gradient. CD4+ T lymphocytes were enriched from total PBMCs by negative sorting. HIV DNA concentration in magnetic beads enriched group and whole blood group was detected by HIV DNA detection kit. Results CD4+ T cells were isolated by magnetic beads and identified by FCM for purity at (96.4 ± 2.6)%. The viability was (95.9 ± 2.9)%, as demonstrated by trypan blue staining. The person on continued ART treatment in this study had significantly greater reduction in HIV viral load and undetectable HIV plasma RNA at follow up timepoint 4. No HIV DNA was detected in the whole blood group at all 4 timepoints. The quantitative results of HIV DNA in the CD4+ T lymphocyte group of the magnetic bead enrichment group were 73.4, 429.3, 137.1, 449.9 copies/106 CD4+ T cell′s respectively. Conclusion The magnetic bead enrichment method can be more sensitive in detecting the limit low copy HIV DNA in blood samples, and provide early confirmatory data for HIV WP infection and breakthrough infection after ART treatment.

Authors and Affiliations

Lilin WANG, Linfeng WU, Fang ZHAO, Ran LI, Tong LI, Heng LIU, Rui ZHU, Jinfeng ZENG

Keywords

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EP ID EP738715
DOI 10.13303/j.cjbt.issn.1004-549x.2023.12.006
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