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Viruses 2016, 8(8), 233;

Quantification of HEV RNA by Droplet Digital PCR

CHU Toulouse, Hôpital Purpan, Laboratoire de virologie, Institut fédératif de biologie, Toulouse F-31300, France
INSERM, U1043, Centre de Physiopathologie de Toulouse Purpan, Toulouse F-31300, France
Université Toulouse III Paul-Sabatier, Faculté de Médecine Toulouse, Toulouse F-31300, France
CHU Toulouse, Hôpital Rangueil, Département de Néphrologie, Dialyse et Transplantation multi-organe, Toulouse F-31300, France
Author to whom correspondence should be addressed.
Academic Editor: Eric O. Freed
Received: 17 June 2016 / Revised: 13 August 2016 / Accepted: 16 August 2016 / Published: 19 August 2016
(This article belongs to the Special Issue Recent Progress in Hepatitis E Virus Research)
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The sensitivity of real-time PCR for hepatitis E virus (HEV) RNA quantification differs greatly among techniques. Standardized tools that measure the real quantity of virus are needed. We assessed the performance of a reverse transcription droplet digital PCR (RT-ddPCR) assay that gives absolute quantities of HEV RNA. Analytical and clinical validation was done on HEV genotypes 1, 3 and 4, and was based on open reading frame (ORF)3 amplification. The within-run and between-run reproducibilities were very good, the analytical sensitivity was 80 HEV RNA international units (IU)/mL and linearities of HEV genotype 1, 3 and 4 were very similar. Clinical validation based on 45 samples of genotype 1, 3 or 4 gave results that correlated well with a validated reverse transcription quantitative PCR (RT-qPCR) assay (Spearman rs = 0.89, p < 0.0001). The RT-ddPCR assay is a sensitive method and could be a promising tool for standardizing HEV RNA quantification in various sample types. View Full-Text
Keywords: ddPCR; absolute quantification; HEV quantification ddPCR; absolute quantification; HEV quantification

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Nicot, F.; Cazabat, M.; Lhomme, S.; Marion, O.; Sauné, K.; Chiabrando, J.; Dubois, M.; Kamar, N.; Abravanel, F.; Izopet, J. Quantification of HEV RNA by Droplet Digital PCR. Viruses 2016, 8, 233.

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