Quantification of HEV RNA by Droplet Digital PCR
AbstractThe 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
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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.
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(8):233.Chicago/Turabian Style
Nicot, Florence; Cazabat, Michelle; Lhomme, Sébastien; Marion, Olivier; Sauné, Karine; Chiabrando, Julie; Dubois, Martine; Kamar, Nassim; Abravanel, Florence; Izopet, Jacques. 2016. "Quantification of HEV RNA by Droplet Digital PCR." Viruses 8, no. 8: 233.
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