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Article: Rapid quantification of hepatitis B virus DNA by real-time PCR using fluorescent hybridization probes

TitleRapid quantification of hepatitis B virus DNA by real-time PCR using fluorescent hybridization probes
Authors
Issue Date2003
PublisherSociety for General Microbiology. The Journal's web site is located at http://jmm.sgmjournals.org
Citation
Journal Of Medical Microbiology, 2003, v. 52 n. 5, p. 397-402 How to Cite?
AbstractA highly sensitive and rapid assay has been developed to quantify hepatitis B virus (HBV) DNA, based on the fluorescence resonance energy transfer principle and real-time PCR, using the LightCycler and a pair of specific fluorescent hybridization probes. This LightCycler real-time PCR assay (LC-PCR) detected HBV DNA in a linear range from 10 1 to 10 8 copies per reaction (250-2.5 × 10 9 copies ml -1), with a rapid PCR cycling time of 35 min. The assay was validated with two EUROHEP HBV DNA standards (ad and ay subtypes) and exhibited low intra-assay (< 6%) and inter-assay (< 16%) variation for both subtypes over the complete range of 7 orders of magnitude. The assay was evaluated clinically using serum samples from 120 HBsAg + individuals and 45 healthy controls who were negative for both HBsAg and anti-HBc. Levels of HBV DNA were measured in these samples using both the LC-PCR and Digene Hybrid Capture II HBV DNA (HCII) assays. The prevalence rates for HBV DNA in the HBsAg + serum samples were respectively 95% (114/120) and 56% (67/120) by LC-PCR and HCII (P < 0.01). All 67 HCII-positive samples tested positive with LC-PCR, while the 47 discordant samples showed low levels of HBV DNA (down to 265 copies ml -1), detectable only by the more sensitive LC-PCR assay. Levels of HBV DNA as measured by the two assays showed good correlation (r = 0.902; P < 0.001). The level of HBV DNA was significantly higher in HBeAg + than anti-HBe + samples (median 1.5 × 10 7 vs 4.6 × 10 4 copies ml -1; P < 0.01). It is concluded that this LC-PCR assay is clinically useful for the rapid, sensitive and accurate measurement of HBV DNA.
Persistent Identifierhttp://hdl.handle.net/10722/77928
ISSN
2015 Impact Factor: 2.269
2015 SCImago Journal Rankings: 1.060
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorHo, SKNen_HK
dc.contributor.authorYam, WCen_HK
dc.contributor.authorLeung, ETKen_HK
dc.contributor.authorWong, LPen_HK
dc.contributor.authorLeung, JKHen_HK
dc.contributor.authorLai, KNen_HK
dc.contributor.authorChan, TMen_HK
dc.date.accessioned2010-09-06T07:37:17Z-
dc.date.available2010-09-06T07:37:17Z-
dc.date.issued2003en_HK
dc.identifier.citationJournal Of Medical Microbiology, 2003, v. 52 n. 5, p. 397-402en_HK
dc.identifier.issn0022-2615en_HK
dc.identifier.urihttp://hdl.handle.net/10722/77928-
dc.description.abstractA highly sensitive and rapid assay has been developed to quantify hepatitis B virus (HBV) DNA, based on the fluorescence resonance energy transfer principle and real-time PCR, using the LightCycler and a pair of specific fluorescent hybridization probes. This LightCycler real-time PCR assay (LC-PCR) detected HBV DNA in a linear range from 10 1 to 10 8 copies per reaction (250-2.5 × 10 9 copies ml -1), with a rapid PCR cycling time of 35 min. The assay was validated with two EUROHEP HBV DNA standards (ad and ay subtypes) and exhibited low intra-assay (< 6%) and inter-assay (< 16%) variation for both subtypes over the complete range of 7 orders of magnitude. The assay was evaluated clinically using serum samples from 120 HBsAg + individuals and 45 healthy controls who were negative for both HBsAg and anti-HBc. Levels of HBV DNA were measured in these samples using both the LC-PCR and Digene Hybrid Capture II HBV DNA (HCII) assays. The prevalence rates for HBV DNA in the HBsAg + serum samples were respectively 95% (114/120) and 56% (67/120) by LC-PCR and HCII (P < 0.01). All 67 HCII-positive samples tested positive with LC-PCR, while the 47 discordant samples showed low levels of HBV DNA (down to 265 copies ml -1), detectable only by the more sensitive LC-PCR assay. Levels of HBV DNA as measured by the two assays showed good correlation (r = 0.902; P < 0.001). The level of HBV DNA was significantly higher in HBeAg + than anti-HBe + samples (median 1.5 × 10 7 vs 4.6 × 10 4 copies ml -1; P < 0.01). It is concluded that this LC-PCR assay is clinically useful for the rapid, sensitive and accurate measurement of HBV DNA.en_HK
dc.languageengen_HK
dc.publisherSociety for General Microbiology. The Journal's web site is located at http://jmm.sgmjournals.orgen_HK
dc.relation.ispartofJournal of Medical Microbiologyen_HK
dc.subject.meshCarrier State - diagnosis - virologyen_HK
dc.subject.meshCase-Control Studiesen_HK
dc.subject.meshDNA Probes - diagnostic useen_HK
dc.subject.meshDNA, Viral - blooden_HK
dc.subject.meshHepatitis B - diagnosis - virologyen_HK
dc.subject.meshHepatitis B Surface Antigens - blooden_HK
dc.subject.meshHepatitis B e Antigens - blooden_HK
dc.subject.meshHepatitis B virus - genetics - isolation & purificationen_HK
dc.subject.meshHumansen_HK
dc.subject.meshIn Situ Hybridization, Fluorescenceen_HK
dc.subject.meshPolymerase Chain Reaction - methodsen_HK
dc.subject.meshReproducibility of Resultsen_HK
dc.subject.meshSensitivity and Specificityen_HK
dc.subject.meshTime Factorsen_HK
dc.subject.meshViral Loaden_HK
dc.titleRapid quantification of hepatitis B virus DNA by real-time PCR using fluorescent hybridization probesen_HK
dc.typeArticleen_HK
dc.identifier.openurlhttp://library.hku.hk:4550/resserv?sid=HKU:IR&issn=0022-2615&volume=52&spage=397&epage=402&date=2003&atitle=Rapid+quantification+of+hepatitis+B+virus+DNA+by+real-time+PCR+using+fluorescent+hybridization+probesen_HK
dc.identifier.emailYam, WC: wcyam@hkucc.hku.hken_HK
dc.identifier.emailLai, KN: knlai@hku.hken_HK
dc.identifier.emailChan, TM: dtmchan@hku.hken_HK
dc.identifier.authorityYam, WC=rp00313en_HK
dc.identifier.authorityLai, KN=rp00324en_HK
dc.identifier.authorityChan, TM=rp00394en_HK
dc.description.naturelink_to_subscribed_fulltext-
dc.identifier.doi10.1099/jmm.0.05071-0en_HK
dc.identifier.pmid12721315-
dc.identifier.scopuseid_2-s2.0-0037719633en_HK
dc.identifier.hkuros79062en_HK
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-0037719633&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume52en_HK
dc.identifier.issue5en_HK
dc.identifier.spage397en_HK
dc.identifier.epage402en_HK
dc.identifier.isiWOS:000183011800006-
dc.publisher.placeUnited Kingdomen_HK
dc.identifier.scopusauthoridHo, SKN=36839065300en_HK
dc.identifier.scopusauthoridYam, WC=7004281720en_HK
dc.identifier.scopusauthoridLeung, ETK=36788339100en_HK
dc.identifier.scopusauthoridWong, LP=7402092221en_HK
dc.identifier.scopusauthoridLeung, JKH=7202180353en_HK
dc.identifier.scopusauthoridLai, KN=7402135706en_HK
dc.identifier.scopusauthoridChan, TM=7402687700en_HK
dc.identifier.citeulike4544388-

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