Linked Life Data

10203493

Source:http://linkedlifedata.com/resource/pubmed/id/10203493

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
5
pubmed:dateCreated
1999-5-7
pubmed:databankReference
pubmed:abstractText
The genome of hepatitis C virus (HCV) consists of seven functional regions: the core, E1, E2/NS1, NS2, NS3, NS4, and NS5 regions. The U. S. Food and Drug Administration-licensed 2.0G immunoassay for the detection of anti-HCV uses proteins from the core, NS3, and NS4 regions (McHutchinson et al., Hepatology 15:19-25, 1992). The 3.0G enzyme-linked immunosorbent assay includes the protein from the NS5 region (Uyttendaele et al., Vox Sang. 66:122-129, 1994). The necessity of detecting antibodies to viral envelope proteins (E1 and E2) and to different genotype samples has been demonstrated previously (Chien et al., Lancet 342:933, 1993; Lok et al., Hepatology 18:497-502, 1993). In this study we have attempted to improve the sensitivity of the anti-HCV assay by developing a single multiple-epitope fusion antigen (MEFA; MEFA-6) which incorporates all of the major immunodominant epitopes from the seven functional regions of the HCV genome. A nucleic acid sequence consisting of proteins from the viral core, E1, E2, NS3, NS4, and NS5 regions and different subtype-specific regions of the NS4 region was constructed, cloned, and expressed in yeast. The epitopes present on this antigen can be detected by epitope-specific monoclonal and polyclonal antibodies. In a competition assay, the MEFA-6 protein competed with 83 to 96% of genotype-specific antibodies from HCV genotype-specific peptides. This recombinant antigen was subsequently used to design an anti-HCV chemiluminescent immunoassay. We designed our assay using a monoclonal anti-human immunoglobulin G antibody bound to the solid phase. Because MEFA-6 is fused with human superoxide dismutase (h-SOD), we used an anti-human superoxide dismutase, dimethyl acridinium ester-labeled monoclonal antibody for detection. Our results indicate that MEFA-6 exposes all of the major immunogenic epitopes. Its excellent sensitivity and specificity for the detection of clinical seroconversion are demonstrated by this assay.
pubmed:commentsCorrections
http://linkedlifedata.com/resource/pubmed/commentcorrection/10203493-1279666, http://linkedlifedata.com/resource/pubmed/commentcorrection/10203493-1309365, http://linkedlifedata.com/resource/pubmed/commentcorrection/10203493-1373489, http://linkedlifedata.com/resource/pubmed/commentcorrection/10203493-1656258, http://linkedlifedata.com/resource/pubmed/commentcorrection/10203493-1848704, http://linkedlifedata.com/resource/pubmed/commentcorrection/10203493-2496467, http://linkedlifedata.com/resource/pubmed/commentcorrection/10203493-2523562, http://linkedlifedata.com/resource/pubmed/commentcorrection/10203493-2647749, http://linkedlifedata.com/resource/pubmed/commentcorrection/10203493-3889846, http://linkedlifedata.com/resource/pubmed/commentcorrection/10203493-7514324, http://linkedlifedata.com/resource/pubmed/commentcorrection/10203493-7686182, http://linkedlifedata.com/resource/pubmed/commentcorrection/10203493-7689528, http://linkedlifedata.com/resource/pubmed/commentcorrection/10203493-7692197
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
May
pubmed:issn
0095-1137
pubmed:author
pubmed:issnType
Print
pubmed:volume
37
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
1393-7
pubmed:dateRevised
2009-11-18
pubmed:meshHeading
pubmed:year
1999
pubmed:articleTitle
Use of a novel hepatitis C virus (HCV) major-epitope chimeric polypeptide for diagnosis of HCV infection.
pubmed:affiliation
Chiron Corporation, Emeryville, California 94507, USA. david_chien@cc.chiron.com
pubmed:publicationType
Journal Article