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J Clin Virol. 2015 Apr;65:38-40. doi: 10.1016/j.jcv.2015.01.022. Epub 2015 Feb 1.

A simple method to elute cell-free HIV from dried blood spots improves their usefulness for monitoring therapy.

Author information

1
Roche Molecular Systems, Pleasanton, CA, USA.
2
Roche Molecular Systems, Pleasanton, CA, USA. Electronic address: paul.baum@roche.com.

Abstract

BACKGROUND:

Dried blood spots (DBS) improve access to HIV viral load (VL) testing, but yield increased VL measurements compared to the plasma reference method because of cell-associated viral nucleic acid. In clinical settings, DBS methods may falsely categorize many patients as failing therapy.

OBJECTIVES:

Description of a simple method, free virus elution (FVE), to preferentially elute plasma-associated virus from DBS samples with phosphate-buffered saline, and an initial HIV VL performance comparison with standard DBS elution methods.

STUDY DESIGN:

The mechanism of action of FVE was studied with model DBS samples containing purified virus or washed HIV-containing cells, and with a DNA-specific HIV PCR. Using clinical samples, VL results from the new FVE method were compared to results from a dried fluid spot procedure (DFSP) protocol, which uses a guanidinium-based elution method, using plasma VL as the reference method.

RESULTS:

Model system experiments suggest that the method efficiently separates virus from cell-associated HIV, with a wide tolerance for incubation time and temperature. In 196 clinical samples, FVE reduced VL over-quantification from DBS, and improved DBS clinical concordance with plasma from 67% to 95%.

CONCLUSIONS:

A simple elution in PBS significantly reduced the over-quantification of HIV VL in DBS. Additional studies are needed to validate the method in fingerstick-collected specimens and to further understand the compartmentalization of HIV DNA and RNA in DBS specimens.

KEYWORDS:

DBS; Dried blood spots; HIV; Viral load

PMID:
25766985
DOI:
10.1016/j.jcv.2015.01.022
[Indexed for MEDLINE]
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