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Detection and quantification of hepatitis B virus DNA by SYBR green real-time polymerase chain reaction.
- Source :
-
European journal of clinical microbiology & infectious diseases : official publication of the European Society of Clinical Microbiology [Eur J Clin Microbiol Infect Dis] 2007 Jan; Vol. 26 (1), pp. 43-50. - Publication Year :
- 2007
-
Abstract
- A single-round real-time 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 real-time PCR assay was evaluated in a routine clinical laboratory setting with a total of 212 clinical specimens. The sensitivity of the real-time 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 10(3)-10(9) IU/ml (2.28 x 10(3) to 2.28 x 10(9) copies/ml), was observed in the quantitative determination. The interexperimental variability coefficient of the assay ranged from 0.22 to 0.39 and the intraexperimental variability coefficient from 0.24 to 0.41. By excluding values outside of the dynamic ranges of both tests, the HBV Monitor and the real-time 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 1.5 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 real-time 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.
- Subjects :
- Benzothiazoles
Diamines
Hepatitis B, Chronic virology
Humans
Molecular Sequence Data
Organic Chemicals
Polymerase Chain Reaction
Quinolines
Sensitivity and Specificity
Viral Load methods
DNA, Viral analysis
Hepatitis B virus genetics
Hepatitis B, Chronic diagnosis
Reagent Kits, Diagnostic virology
Subjects
Details
- Language :
- English
- ISSN :
- 0934-9723
- Volume :
- 26
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- European journal of clinical microbiology & infectious diseases : official publication of the European Society of Clinical Microbiology
- Publication Type :
- Academic Journal
- Accession number :
- 17216291
- Full Text :
- https://doi.org/10.1007/s10096-006-0223-y