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Article: Micromethod for phosphonoformate inhibition assay of hepatitis B viral DNA polymerase.

TitleMicromethod for phosphonoformate inhibition assay of hepatitis B viral DNA polymerase.
Authors
Issue Date1984
PublisherAmerican Association for Clinical Chemistry, Inc. The Journal's web site is located at http://www.clinchem.org
Citation
Clinical Chemistry, 1984, v. 30 n. 4, p. 549-552 How to Cite?
AbstractA micromethod for the specific measurement of hepatitis B viral DNA polymerase in serum is presented, based on the phosphonoformate inhibition assay (J Med Virol 12: 61-70, 1983). In the micromethod, sample volume is reduced to 120 microL and the ultracentrifugation step is eliminated. The method allows good discrimination between serum infected with hepatitis B virus and uninfected serum. The cutoff value for rate of nucleotide incorporation, based on assays of 41 serum specimens negative for hepatitis B serological markers, was about 15 nU/L (90th percentile). Serum containing hepatitis B surface and antigens exhibited rates of phosphonoformate-inhibitive nucleotide incorporation of 150 (SD 150) nU/L, with an upper 90th percentile range of 17 to 667 nU/L (n = 41). The micromethod makes use of commercially available [32P]dCTP (specific activity about 7000 kCi/mol). 125I-labeled dCTP was found to be unsuitable for this assay. Human DNA polymerases in serum are detected by this method but are excluded from the phosphonoformate-inhibitive fraction.
Persistent Identifierhttp://hdl.handle.net/10722/161679
ISSN
2015 Impact Factor: 7.457
2015 SCImago Journal Rankings: 2.472
ISI Accession Number ID

 

DC FieldValueLanguage
dc.contributor.authorLin, HJen_US
dc.contributor.authorWu, PCen_US
dc.contributor.authorLai, CLen_US
dc.contributor.authorChak, Wen_US
dc.date.accessioned2012-09-05T05:13:51Z-
dc.date.available2012-09-05T05:13:51Z-
dc.date.issued1984en_US
dc.identifier.citationClinical Chemistry, 1984, v. 30 n. 4, p. 549-552en_US
dc.identifier.issn0009-9147en_US
dc.identifier.urihttp://hdl.handle.net/10722/161679-
dc.description.abstractA micromethod for the specific measurement of hepatitis B viral DNA polymerase in serum is presented, based on the phosphonoformate inhibition assay (J Med Virol 12: 61-70, 1983). In the micromethod, sample volume is reduced to 120 microL and the ultracentrifugation step is eliminated. The method allows good discrimination between serum infected with hepatitis B virus and uninfected serum. The cutoff value for rate of nucleotide incorporation, based on assays of 41 serum specimens negative for hepatitis B serological markers, was about 15 nU/L (90th percentile). Serum containing hepatitis B surface and antigens exhibited rates of phosphonoformate-inhibitive nucleotide incorporation of 150 (SD 150) nU/L, with an upper 90th percentile range of 17 to 667 nU/L (n = 41). The micromethod makes use of commercially available [32P]dCTP (specific activity about 7000 kCi/mol). 125I-labeled dCTP was found to be unsuitable for this assay. Human DNA polymerases in serum are detected by this method but are excluded from the phosphonoformate-inhibitive fraction.en_US
dc.languageengen_US
dc.publisherAmerican Association for Clinical Chemistry, Inc. The Journal's web site is located at http://www.clinchem.orgen_US
dc.relation.ispartofClinical Chemistryen_US
dc.subject.meshDna-Directed Dna Polymerase - Antagonists & Inhibitors - Blooden_US
dc.subject.meshDeoxycytosine Nucleotides - Metabolismen_US
dc.subject.meshFoscarneten_US
dc.subject.meshHepatitis B - Enzymologyen_US
dc.subject.meshHepatitis B Virus - Enzymologyen_US
dc.subject.meshHumansen_US
dc.subject.meshKineticsen_US
dc.subject.meshMicrochemistry - Methodsen_US
dc.subject.meshOrganophosphorus Compounds - Pharmacologyen_US
dc.subject.meshPhosphonoacetic Acid - Analogs & Derivatives - Pharmacologyen_US
dc.titleMicromethod for phosphonoformate inhibition assay of hepatitis B viral DNA polymerase.en_US
dc.typeArticleen_US
dc.identifier.emailLai, CL:hrmelcl@hku.hken_US
dc.identifier.authorityLai, CL=rp00314en_US
dc.description.naturelink_to_subscribed_fulltexten_US
dc.identifier.pmid6231137-
dc.identifier.scopuseid_2-s2.0-0021414186en_US
dc.identifier.volume30en_US
dc.identifier.issue4en_US
dc.identifier.spage549en_US
dc.identifier.epage552en_US
dc.identifier.isiWOS:A1984SL74200011-
dc.publisher.placeUnited Statesen_US
dc.identifier.scopusauthoridLin, HJ=7405571292en_US
dc.identifier.scopusauthoridWu, PC=7403119323en_US
dc.identifier.scopusauthoridLai, CL=7403086396en_US
dc.identifier.scopusauthoridChak, W=6602179224en_US

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