Development of a new ultra sensitive real-time PCR assay (ultra sensitive RTQ-PCR) for the quantification of HBV-DNA

Paraskevis, Dimitrios, Beloukas, Apostolos ORCID: 0000-0001-5639-0528, Haida, Catherine, Katsoulidou, Antigoni, Moschidis, Zisis, Hatzitheodorou, Helen, Varaklioti, Agoritsa, Sypsa, Vana and Hatzakis, Angelos (2010) Development of a new ultra sensitive real-time PCR assay (ultra sensitive RTQ-PCR) for the quantification of HBV-DNA. VIROLOGY JOURNAL, 7 (1). 57-.

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Abstract

<h4>Background</h4>Improved sensitivity of HBV-DNA tests is of critical importance for the management of HBV infection. Our aim was to develop and assess a new ultra sensitive in-house real-time PCR assay for HBV-DNA quantification (ultra sensitive RTQ-PCR).<h4>Results</h4>Previously used HBV-DNA standards were calibrated against the WHO 1st International Standard for HBV-DNA (OptiQuant(R) HBV-DNA Quantification Panel, Accrometrix Europe B.V.). The 95% and 50% HBV-DNA detection end-point of the assay were 22.2 and 8.4 IU/mL. According to the calibration results, 1 IU/mL equals 2.8 copies/mL. Importantly the clinical performance of the ultra sensitive real-time PCR was tested similar (67%) to the Procleix Ultrio discriminatory HBV test (dHBV) (70%) in low-titer samples from patients with occult Hepatitis B. Finally, in the comparison of ultra sensitive RTQ-PCR with the commercially available COBAS TaqMan HBV Test, the in-house assay identified 94.7% of the 94 specimens as positive versus 90.4% identified by TaqMan, while the quantitative results that were positive by both assay were strongly correlated (r = 0.979).<h4>Conclusions</h4>We report a new ultra sensitive real time PCR molecular beacon based assay with remarkable analytical and clinical sensitivity, calibrated against the WHO 1st International standard.

Item Type:	Article
Uncontrolled Keywords:	Humans, Hepatitis B virus, Hepatitis B, DNA, Viral, Viral Load, Sensitivity and Specificity, Polymerase Chain Reaction
Depositing User:	Symplectic Admin
Date Deposited:	19 Oct 2015 14:34
Last Modified:	16 Dec 2022 03:00
DOI:	10.1186/1743-422X-7-57
Publisher's Statement :	© 2010 Paraskevis et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Related URLs:	Author Publisher
URI:	https://livrepository.liverpool.ac.uk/id/eprint/2032401