Τίτλος:
Development of a new ultra sensitive real-time PCR assay (ultra sensitive RTQ-PCR) for the quantification of HBV-DNA
Γλώσσες Τεκμηρίου:
Αγγλικά
Περίληψη:
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 1st 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 1st International standard. © 2010 Paraskevis et al; licensee BioMed Central Ltd.
Συγγραφείς:
Paraskevis, D.
Beloukas, A.
Haida, C.
Katsoulidou, A.
Moschidis, Z.
Hatzitheodorou, H.
Varaklioti, A.
Sypsa, V.
Hatzakis, A.
Περιοδικό:
Virology Journal
Λέξεις-κλειδιά:
DNA; virus DNA, article; controlled study; hepatitis B; Hepatitis B virus; intermethod comparison; nonhuman; performance measurement system; real time polymerase chain reaction; sensitivity analysis; virus detection; blood; comparative study; evaluation; genetics; hepatitis B; human; isolation and purification; methodology; polymerase chain reaction; sensitivity and specificity; virus load, Hepatitis B virus, DNA, Viral; Hepatitis B; Hepatitis B virus; Humans; Polymerase Chain Reaction; Sensitivity and Specificity; Viral Load
DOI:
10.1186/1743-422X-7-57
