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PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
2
pubmed:dateCreated
2004-3-11
pubmed:abstractText
To identify a major antigenic determinant for use in the development of a rapid serological diagnostic test for severe acute respiratory syndrome (SARS) coronavirus infection and to study the immune response during SARS coronavirus infection in humans, we cloned the full length and six truncated fragments of the nucleocapsid gene, expressed them, and purified them as glutathione S-transferase-tagged recombinant proteins. The reactivities of the recombinant proteins to a panel of antibodies containing 33 SARS coronavirus-positive sera and 66 negative sera and to antibodies against other animal coronaviruses were screened. A truncated 195-amino-acid fragment from the C terminus of the nucleocapsid protein (N195) was identified that had a strong ability to detect antibodies against SARS coronavirus. No cross-reaction was found between the N195 protein and antibodies against chicken, pig, and canine coronaviruses. The N195 protein was used to develop a Western blot assay to detect antibodies against SARS coronavirus in 274 clinically blinded samples. The specificity and sensitivity of this test were 98.3 and 90.9%, respectively. The correlation between our Western blotting assay and an immunofluorescence assay (IFA) was also analyzed. The results of our Western blot assay and IFA for the detection of SARS coronavirus-positive sera were the same. Thus, the N195 protein was identified as a suitable protein to be used as an antigen in Western blot and other possible assays for the detection of SARS coronavirus infection.
pubmed:commentsCorrections
http://linkedlifedata.com/resource/pubmed/commentcorrection/15013997-10458108, http://linkedlifedata.com/resource/pubmed/commentcorrection/15013997-10865148, http://linkedlifedata.com/resource/pubmed/commentcorrection/15013997-12671061, http://linkedlifedata.com/resource/pubmed/commentcorrection/15013997-12690091, http://linkedlifedata.com/resource/pubmed/commentcorrection/15013997-12690092, http://linkedlifedata.com/resource/pubmed/commentcorrection/15013997-12711465, http://linkedlifedata.com/resource/pubmed/commentcorrection/15013997-12730500, http://linkedlifedata.com/resource/pubmed/commentcorrection/15013997-12730501, http://linkedlifedata.com/resource/pubmed/commentcorrection/15013997-12781535, http://linkedlifedata.com/resource/pubmed/commentcorrection/15013997-12781537, http://linkedlifedata.com/resource/pubmed/commentcorrection/15013997-12791839, http://linkedlifedata.com/resource/pubmed/commentcorrection/15013997-12800213, http://linkedlifedata.com/resource/pubmed/commentcorrection/15013997-12865207, http://linkedlifedata.com/resource/pubmed/commentcorrection/15013997-12890855, http://linkedlifedata.com/resource/pubmed/commentcorrection/15013997-1310335, http://linkedlifedata.com/resource/pubmed/commentcorrection/15013997-7516595, http://linkedlifedata.com/resource/pubmed/commentcorrection/15013997-9079758
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Mar
pubmed:issn
1071-412X
pubmed:author
pubmed:issnType
Print
pubmed:volume
11
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
417-22
pubmed:dateRevised
2009-11-18
pubmed:meshHeading
pubmed:year
2004
pubmed:articleTitle
Development of a Western blot assay for detection of antibodies against coronavirus causing severe acute respiratory syndrome.
pubmed:affiliation
Animal Health Biotechnology, Temasek Life Science Laboratory, National University of Singapore, Republic of Singapore.
pubmed:publicationType
Journal Article