Immiscible phase filter extraction and equivalent amplification of genotypes 1–6 of hepatitis C RNA: The building blocks for point-of-care diagnosis

Mário F. Neto, Matthew A. Butzler, Jennifer L. Reed, Xiang Rui, Mark J Fisher, David M Kelso, Sally M McFall*

*Corresponding author for this work

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

The lack of hepatitis C virus (HCV) diagnostic tests designed for use in decentralized settings is a major obstacle for providing access to treatment and prevention services particularly in low and middle income countries. Here we describe the development and validation of two building blocks of the HCV Quant Assay, a test in development for point-of-care use: 1) an RT-qPCR assay with noncompetitive internal control that equivalently detects the 6 major HCV genotypes and 2) an automated sample prep method using immiscible phase filter technology. This novel assay has wide dynamic range of HCV quantification and a limit of detection of 30 IU/ml with 200 μl specimen volume. In a preliminary study of 61 clinical specimens, the HCV Quant Assay demonstrated 100% sensitivity and specificity and gave comparable viral load results across 4 logs of IU/ml when compared to the Abbott RealTime HCV Assay.

Original languageEnglish (US)
Pages (from-to)107-115
Number of pages9
JournalJournal of Virological Methods
Volume248
DOIs
StatePublished - Oct 1 2017

Fingerprint

Point-of-Care Systems
Hepatitis C
Hepacivirus
Genotype
RNA
Viral Load
Routine Diagnostic Tests
Limit of Detection
Technology
Sensitivity and Specificity

Keywords

  • HCV
  • RT-qPCR
  • Sample prep
  • Viral load monitoring

ASJC Scopus subject areas

  • Virology

Cite this

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title = "Immiscible phase filter extraction and equivalent amplification of genotypes 1–6 of hepatitis C RNA: The building blocks for point-of-care diagnosis",
abstract = "The lack of hepatitis C virus (HCV) diagnostic tests designed for use in decentralized settings is a major obstacle for providing access to treatment and prevention services particularly in low and middle income countries. Here we describe the development and validation of two building blocks of the HCV Quant Assay, a test in development for point-of-care use: 1) an RT-qPCR assay with noncompetitive internal control that equivalently detects the 6 major HCV genotypes and 2) an automated sample prep method using immiscible phase filter technology. This novel assay has wide dynamic range of HCV quantification and a limit of detection of 30 IU/ml with 200 μl specimen volume. In a preliminary study of 61 clinical specimens, the HCV Quant Assay demonstrated 100{\%} sensitivity and specificity and gave comparable viral load results across 4 logs of IU/ml when compared to the Abbott RealTime HCV Assay.",
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Immiscible phase filter extraction and equivalent amplification of genotypes 1–6 of hepatitis C RNA : The building blocks for point-of-care diagnosis. / Neto, Mário F.; Butzler, Matthew A.; Reed, Jennifer L.; Rui, Xiang; Fisher, Mark J; Kelso, David M; McFall, Sally M.

In: Journal of Virological Methods, Vol. 248, 01.10.2017, p. 107-115.

Research output: Contribution to journalArticle

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AU - Neto, Mário F.

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AU - Rui, Xiang

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AU - Kelso, David M

AU - McFall, Sally M

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