TY - JOUR
TI - Development of a new ultra sensitive real-time PCR assay (ultra sensitive RTQ-PCR) for the quantification of HBV-DNA
AU - Paraskevis, D.
AU - Beloukas, A.
AU - Haida, C.
AU - Katsoulidou, A.
AU - Moschidis, Z.
AU - Hatzitheodorou, H.
AU - Varaklioti, A.
AU - Sypsa, V.
AU - Hatzakis, A.
JO - Virology Journal
PY - 2010
VL - 7
TODO - null
SP - null
PB - 
SN - 1743-422X
TODO - 10.1186/1743-422X-7-57
TODO - 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
TODO - 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.
ER -