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

BACKGROUND: 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). RESULTS: Previously used HBV-DNA standards were...

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Autores principales: Paraskevis, Dimitrios, Beloukas, Apostolos, Haida, Catherine, Katsoulidou, Antigoni, Moschidis, Zisis, Hatzitheodorou, Helen, Varaklioti, Agoritsa, Sypsa, Vana, Hatzakis, Angelos
Formato: Texto
Lenguaje:English
Publicado: BioMed Central 2010
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2848214/
https://www.ncbi.nlm.nih.gov/pubmed/20226057
http://dx.doi.org/10.1186/1743-422X-7-57
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author Paraskevis, Dimitrios
Beloukas, Apostolos
Haida, Catherine
Katsoulidou, Antigoni
Moschidis, Zisis
Hatzitheodorou, Helen
Varaklioti, Agoritsa
Sypsa, Vana
Hatzakis, Angelos
author_facet Paraskevis, Dimitrios
Beloukas, Apostolos
Haida, Catherine
Katsoulidou, Antigoni
Moschidis, Zisis
Hatzitheodorou, Helen
Varaklioti, Agoritsa
Sypsa, Vana
Hatzakis, Angelos
author_sort Paraskevis, Dimitrios
collection PubMed
description BACKGROUND: 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). RESULTS: Previously used HBV-DNA standards were calibrated against the WHO 1(st )International Standard for HBV-DNA (OptiQuant(® )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). CONCLUSIONS: We report a new ultra sensitive real time PCR molecular beacon based assay with remarkable analytical and clinical sensitivity, calibrated against the WHO 1(st )International standard.
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spelling pubmed-28482142010-04-01 Development of a new ultra sensitive real-time PCR assay (ultra sensitive RTQ-PCR) for the quantification of HBV-DNA Paraskevis, Dimitrios Beloukas, Apostolos Haida, Catherine Katsoulidou, Antigoni Moschidis, Zisis Hatzitheodorou, Helen Varaklioti, Agoritsa Sypsa, Vana Hatzakis, Angelos Virol J Methodology BACKGROUND: 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). RESULTS: Previously used HBV-DNA standards were calibrated against the WHO 1(st )International Standard for HBV-DNA (OptiQuant(® )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). CONCLUSIONS: We report a new ultra sensitive real time PCR molecular beacon based assay with remarkable analytical and clinical sensitivity, calibrated against the WHO 1(st )International standard. BioMed Central 2010-03-12 /pmc/articles/PMC2848214/ /pubmed/20226057 http://dx.doi.org/10.1186/1743-422X-7-57 Text en Copyright ©2010 Paraskevis et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 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.
spellingShingle Methodology
Paraskevis, Dimitrios
Beloukas, Apostolos
Haida, Catherine
Katsoulidou, Antigoni
Moschidis, Zisis
Hatzitheodorou, Helen
Varaklioti, Agoritsa
Sypsa, Vana
Hatzakis, Angelos
Development of a new ultra sensitive real-time PCR assay (ultra sensitive RTQ-PCR) for the quantification of HBV-DNA
title Development of a new ultra sensitive real-time PCR assay (ultra sensitive RTQ-PCR) for the quantification of HBV-DNA
title_full Development of a new ultra sensitive real-time PCR assay (ultra sensitive RTQ-PCR) for the quantification of HBV-DNA
title_fullStr Development of a new ultra sensitive real-time PCR assay (ultra sensitive RTQ-PCR) for the quantification of HBV-DNA
title_full_unstemmed Development of a new ultra sensitive real-time PCR assay (ultra sensitive RTQ-PCR) for the quantification of HBV-DNA
title_short Development of a new ultra sensitive real-time PCR assay (ultra sensitive RTQ-PCR) for the quantification of HBV-DNA
title_sort development of a new ultra sensitive real-time pcr assay (ultra sensitive rtq-pcr) for the quantification of hbv-dna
topic Methodology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2848214/
https://www.ncbi.nlm.nih.gov/pubmed/20226057
http://dx.doi.org/10.1186/1743-422X-7-57
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