Authors: D Olioso M Boaretti M Ligozzi G Lo Cascio R Fontana
Publish Date: 2006/11/25
Volume: 26, Issue: 1, Pages: 43-50
Abstract
A singleround realtime polymerase chain reaction PCR assay based on SYBR green dye technology for the detection and quantification of hepatitis B virus HBV DNA in serum was evaluated and compared with a qualitative nested PCR and the Cobas Amplicor HBV Monitor assay Roche Molecular Diagnostics Milan Italy The performance of the realtime PCR assay was evaluated in a routine clinical laboratory setting with a total of 212 clinical specimens The sensitivity of the realtime PCR corresponded to 31 IU/ml 70 copies/ml and comparison with the qualitative nested PCR showed significant concordance for 94 of samples The linear curve over 7 log units spanning 103–109 IU/ml 228 × 103 to 228 × 109 copies/ml was observed in the quantitative determination The interexperimental variability coefficient of the assay ranged from 022 to 039 and the intraexperimental variability coefficient from 024 to 041 By excluding values outside of the dynamic ranges of both tests the HBV Monitor and the realtime PCR gave an agreement within ±1 log unit for 90 of samples while those for the remaining 10 were found to be above 1 log unit but less than 15 log units When the results inside and outside the dynamic range of the HBV Monitor were examined 90 of the results were in agreement In conclusion the realtime PCR based on SYBR green technology proved suitable for routine diagnostic purposes showing good sensitivity high specificity high reproducibility and good linearity over a broad dynamic range of quantification
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