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, 13 (7), e0201629
eCollection

Determination of anti-HCV and Quantification of HCV-RNA and IP-10 From Dried Blood Spots Sent by Regular Mail

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Determination of anti-HCV and Quantification of HCV-RNA and IP-10 From Dried Blood Spots Sent by Regular Mail

Bastian Neesgaard et al. PLoS One.

Abstract

Background: With the introduction of direct acting antivirals, treatment of hepatitis C virus (HCV) in hard-to-reach populations is now feasible. Therefore, new cost-effective and reliable test methods are needed. Determination of HCV antibodies and HCV-RNA from dried blood spots samples could represent one such method. Here we examined whether anti-HCV could be detected-and HCV-RNA quantified-from dried blood spots, sent by regular mail. We also investigated, if IP-10 determined from dried blood spots correlated with fibrosis progression appraised by transient elastography.

Method: Forty chronic HCV infected patients were consecutively enrolled. At baseline and after six months, dried blood spots were prepared from blood collected by venous puncture, dried for 4-6 hours, then stored in gas-impermeable plastic bags with a desiccator, before being sent by regular mail. At each visit, approximately six months apart, paired venous samples was obtained and analyzed for anti-HCV, HCV-RNA and IP-10.

Results: Anti-HCV was found in 66/67 of the dried blood spots. Sixty-six paired samples were available for HCV-RNA analysis. A statistically significant correlation was found between log HCV-RNA concentrations in plasma, and log HCV-RNA obtained from (P < 0.0001, Pearson's R 0.6788, R2 0.4607). HCV-RNA, derived from DBS samples, was lower than the corresponding plasma concentration, reflected by a Bland-Altman bias of 3 with SD of bias ± 0.6472. We found no correlation between IP-10 and fibrosis progression.

Conclusions: We identified anti-HCV in 66/67samples, and quantified IP-10 and HCV-RNA from dried blood spots, dried at room temperature and sent by regular mail. HCV-RNA concentrations from the dried blood spots were lower than corresponding plasma values; a probable result of heparin coated test tubes. We found no correlation between IP-10 and fibrosis progression. Overall, dried blood spots could be a cost-effective and easy-to-use alternative to standard tests for the diagnosis of HCV infections.

Conflict of interest statement

At initial submission we had noted that co-author Morten Ruhwald was registered as inventor on a patent application disclosing IP-10 based liver fibrosis monitoring, using DBS, which could be viewed as a conflict-of-interest. However, this application was dropped prior to submission. Thus, it does not constitute a conflict of interest. This does not alter our adherence to PLOS ONE policies on sharing data and materials.

Figures

Fig 1
Fig 1. Linear regression showing correlation between HCV-RNA Concentrations in plasma and DBS samples.
Fig 2
Fig 2. Bland-Altman plot showing difference and average of samples derived from Log plasma HCV-RNA concentrations and Log DBS HCV-RNA concentrations.
Fig 3
Fig 3. Showing no significant difference in plasma IP-10 levels in samples derived from baseline and at 6 months follow-up.

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Grant support

Bastian Neesgaard received grants from Amagar and Hvidovre Hospital Research Foundation of 45000 Dkr and the Family Hede Nielsen Foundation of 10000 Dkr. The funds provided were used as salary for Bastian Neesgaard, during the year of pre graduate research. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
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