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|Title: ||Hepatitis C Virus (HCV) RNA screening and sequencing using dry plasma spots|
|Authors: ||Phillips, Richard O.|
Geretti, Anna Maria
Appiah, Lambert Tetteh
|Keywords: ||HCV RNA|
Dried plasma spots
|Issue Date: ||Oct-2017|
|Publisher: ||Elsevier B.V.|
|Citation: ||Elsevier B.V. 97 (2017) 18–21 19|
|Abstract: ||Background: 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.
Objective: 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.
Study design: In the validation phase, replicate DPS were prepared with six dilutions (500–10,000 IU/ml) of the
4th 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 6 mm punches; pools comprised two
punches from each of five DPS. Genotypes were determined by Sanger sequencing of HCV core and NS5B.
Results: With the dilution series, sensitivity of HCV RNA detection was ≥2500 IU/ml. Replicate DPS gave intraassay
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.
Discussion: 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.|
|Description: ||An article published by Elsevier B.V.|
|Appears in Collections:||College of Health Sciences|
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