Abstract
The sensitivity, specificity, reproducibility, detection level, and quantification potential of the SHARP Signal System for enzymatic detection of amplified hepatitis B virus (HBV) DNA in clinical samples were evaluated by testing 104 samples in parallel in a SHARP PCR, an in-house HBV PCR, and a dot blot hybridization assay for semiquantification. SHARP PCR showed a sensitivity of 100%, a specificity of 92.3% (resolved, 100%), a reproducibility of 92.3% (all discrepant serum samples involved very low levels of HBV DNA), and a detection level of at least 3.5 pg/ml. Clinically relevant quantification of the amplified products was not feasible.
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- Ferre F. Quantitative or semi-quantitative PCR: reality versus myth. PCR Methods Appl. 1992 Aug;2(1):1–9. doi: 10.1101/gr.2.1.1. [DOI] [PubMed] [Google Scholar]
- Jalava T., Lehtovaara P., Kallio A., Ranki M., Söderlund H. Quantification of hepatitis B virus DNA by competitive amplification and hybridization on microplates. Biotechniques. 1993 Jul;15(1):134–139. [PubMed] [Google Scholar]
- Janssen H. L., Schoenmaker-Weber Y. A., Kruining H., Schalm S. W., Heijtink R. A. Quantitative assessment of hepatitis B virus DNA in chronic hepatitis B: comparison of two solution hybridization assays. J Med Virol. 1993 Aug;40(4):307–312. doi: 10.1002/jmv.1890400408. [DOI] [PubMed] [Google Scholar]
- Kaneko S., Feinstone S. M., Miller R. H. Rapid and sensitive method for the detection of serum hepatitis B virus DNA using the polymerase chain reaction technique. J Clin Microbiol. 1989 Sep;27(9):1930–1933. doi: 10.1128/jcm.27.9.1930-1933.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Keller G. H., Huang D. P., Shih J. W., Manak M. M. Detection of hepatitis B virus DNA in serum by polymerase chain reaction amplification and microtiter sandwich hybridization. J Clin Microbiol. 1990 Jun;28(6):1411–1416. doi: 10.1128/jcm.28.6.1411-1416.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Larzul D., Guigue F., Sninsky J. J., Mack D. H., Bréchot C., Guesdon J. L. Detection of hepatitis B virus sequences in serum by using in vitro enzymatic amplification. J Virol Methods. 1988 Jul;20(3):227–237. doi: 10.1016/0166-0934(88)90126-7. [DOI] [PubMed] [Google Scholar]
- Lehtovaara P., Uusi-Oukari M., Buchert P., Laaksonen M., Bengtström M., Ranki M. Quantitative PCR for hepatitis B virus with colorimetric detection. PCR Methods Appl. 1993 Dec;3(3):169–175. doi: 10.1101/gr.3.3.169. [DOI] [PubMed] [Google Scholar]
- Mantero G., Zonaro A., Albertini A., Bertolo P., Primi D. DNA enzyme immunoassay: general method for detecting products of polymerase chain reaction. Clin Chem. 1991 Mar;37(3):422–429. [PubMed] [Google Scholar]
- Noordhoek G. T., Kolk A. H., Bjune G., Catty D., Dale J. W., Fine P. E., Godfrey-Faussett P., Cho S. N., Shinnick T., Svenson S. B. Sensitivity and specificity of PCR for detection of Mycobacterium tuberculosis: a blind comparison study among seven laboratories. J Clin Microbiol. 1994 Feb;32(2):277–284. doi: 10.1128/jcm.32.2.277-284.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pao C. C., Yao D. S., Lin C. Y., Kao S. M., Tsao K. C., Sun C. F., Liaw Y. F. Serum hepatitis B virus DNA in hepatitis B virus seropositive and seronegative patients with normal liver function. Am J Clin Pathol. 1991 Apr;95(4):591–596. doi: 10.1093/ajcp/95.4.591. [DOI] [PubMed] [Google Scholar]
- Quint W. G., Heijtink R. A., Schirm J., Gerlich W. H., Niesters H. G. Reliability of methods for hepatitis B virus DNA detection. J Clin Microbiol. 1995 Jan;33(1):225–228. doi: 10.1128/jcm.33.1.225-228.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Scotto J., Hadchouel M., Hery C., Yvart J., Tiollais P., Brechot C. Detection of hepatitis B virus DNA in serum by a simple spot hybridization technique: comparison with results for other viral markers. Hepatology. 1983 May-Jun;3(3):279–284. doi: 10.1002/hep.1840030301. [DOI] [PubMed] [Google Scholar]
- Thiers V., Nakajima E., Kremsdorf D., Mack D., Schellekens H., Driss F., Goudeau A., Wands J., Sninsky J., Tiollais P. Transmission of hepatitis B from hepatitis-B-seronegative subjects. Lancet. 1988 Dec 3;2(8623):1273–1276. doi: 10.1016/s0140-6736(88)92891-7. [DOI] [PubMed] [Google Scholar]
- Valentine-Thon E., Steinmann J., Arnold W. Detection of hepatitis B virus DNA in serum with nucleic acid probes labelled with 32P, biotin, alkaline phosphatase or sulphone. Mol Cell Probes. 1991 Aug;5(4):299–305. doi: 10.1016/0890-8508(91)90053-m. [DOI] [PubMed] [Google Scholar]
- Valentine-Thon E., Steinmann J., Arnold W. Evaluation of the commercially available HepProbe kit for detection of hepatitis B virus DNA in serum. J Clin Microbiol. 1990 Jan;28(1):39–42. doi: 10.1128/jcm.28.1.39-42.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Zaaijer H. L., Cuypers H. T., Reesink H. W., Winkel I. N., Gerken G., Lelie P. N. Reliability of polymerase chain reaction for detection of hepatitis C virus. Lancet. 1993 Mar 20;341(8847):722–724. doi: 10.1016/0140-6736(93)90488-3. [DOI] [PubMed] [Google Scholar]
- Zyzik E., Gerlich W. H., Uy A., Köchel H., Thomssen R. Assay of hepatitis B virus genome titers in sera of infected subjects. Eur J Clin Microbiol. 1986 Jun;5(3):330–335. doi: 10.1007/BF02017791. [DOI] [PubMed] [Google Scholar]