Hepatitis C Virus (HCV) RNA screening and sequencing using dry plasma spots



Geretti, Anna Maria ORCID: 0000-0002-3670-6588, King, Simon, Adjei-Asante, Kwabena, Appiah, Lambert Tetteh, Owusu, Dorcas Ohui, Sarfo, Fred Stephen, Chadwick, David, Phillips, Richard Odame and Beloukas, Apostolos ORCID: 0000-0001-5639-0528
(2017) Hepatitis C Virus (HCV) RNA screening and sequencing using dry plasma spots. JOURNAL OF CLINICAL VIROLOGY, 97. pp. 18-21.

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Abstract

<h4>Background</h4>HCV RNA screening of large sample repositories provides data on HCV epidemic patterns that may help guide control policies. In resource-limited settings, shipment of frozen samples to molecular laboratory facilities and testing of individual samples may be prohibitively expensive.<h4>Objective</h4>Our aim was to detect and sequence HCV RNA in a large HIV-positive cohort from Kumasi, Ghana, using pooled and individual dried plasma spots (DPS) produced from samples stored at -80°C.<h4>Study design</h4>In the validation phase, replicate DPS were prepared with six dilutions (500-10,000 IU/ml) of the 4<sup>th</sup> International Standard for HCV and tested in three independent experiments. In the testing phase, DPS prepared with plasma samples from 875 HIV-positive subjects were pooled for screening, followed by testing of individual DPS of positive pools. Input from individual DPS was two 6mm punches; pools comprised two punches from each of five DPS. Genotypes were determined by Sanger sequencing of HCV core and NS5B.<h4>Results</h4>With the dilution series, sensitivity of HCV RNA detection was ≥2500 IU/ml. Replicate DPS gave intra-assay and inter-assay coefficients of variation ≤1.4%. With the stored samples, HCV RNA was detected in 5/175 DPS pools and in one DPS from each positive pool, yielding a HCV RNA prevalence of 5/875 (0.57%; 95% confidence interval 0.07-1.07%). The five samples were sequenced as HCV genotypes 2l and 2r.<h4>Discussion</h4>DPS allowed reproducible HCV RNA detection, and pooling effectively contained the cost and labour of screening a previously untested, low-prevalence cohort. DPS were also suitable for HCV sequencing.

Item Type: Article
Uncontrolled Keywords: HCV RNA, Sequencing, Dried plasma spots, Sub-Saharan Africa, Epidemiology
Depositing User: Symplectic Admin
Date Deposited: 03 Nov 2017 12:48
Last Modified: 19 Jan 2023 06:51
DOI: 10.1016/j.jcv.2017.10.012
Open Access URL: http://www.journalofclinicalvirology.com/article/S...
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URI: https://livrepository.liverpool.ac.uk/id/eprint/3011431