Tuyển tập báo cáo các nghiên cứu khoa học quốc tế ngành y học dành cho các bạn tham khảo đề tài: Development of a new ultra sensitive real-time PCR assay (ultra sensitive RTQ-PCR) for the quantification of HBV-DNA | Paraskevis et al. Virology Journal 2010 7 57 http content 7 1 57 VIROLOGY JOURNAL METHODOLOGY Open Access Development of a new ultra sensitive real-time PCR assay ultra sensitive RTQ-PCR for the quantification of HBV-DNA Dimitrios Paraskevis1 Apostolos Beloukas1 Catherine Haida1 Antigoni Katsoulidou1 Zisis Moschidis1 Helen Hatzitheodorou1 Agoritsa Varaklioti2 Vana Sypsa1 Angelos Hatzakis1 Abstract 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 . . The 95 and 50 HBV-DNA detection end-point of the assay were and IU mL. According to the calibration results 1 IU mL equals 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 of the 94 specimens as positive versus identified by TaqMan while the quantitative results that were positive by both assay were strongly correlated r . 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. Background Chronic hepatitis B virus HBV infection can be assessed by evaluating clinical features and biochemical virologic and histologic parameters. Knowledge of HBV-DNA viral load is useful for predicting disease prognosis determining infectivity evaluating indications for treatment .
