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Titolo:
High level expression of recombinant mumps nucleoprotein in Saccharomyces cerevisiae and its evaluation in mumps IgM serology
Autore:
Samuel, D; Sasnauskas, K; Jin, L; Beard, S; Zvirbliene, A; Gedvilaite, A; Cohen, B;
Indirizzi:
Cent Publ Hlth Lab, Enter Resp & Neurol Virus Lab, London NW9 5HT, EnglandCent Publ Hlth Lab London England NW9 5HT s Lab, London NW9 5HT, England Inst Biotechnol, Vilnius, Lithuania Inst Biotechnol Vilnius LithuaniaInst Biotechnol, Vilnius, Lithuania
Titolo Testata:
JOURNAL OF MEDICAL VIROLOGY
fascicolo: 1, volume: 66, anno: 2002,
pagine: 123 - 130
SICI:
0146-6615(200201)66:1<123:HLEORM>2.0.ZU;2-#
Fonte:
ISI
Lingua:
ENG
Soggetto:
ENZYME-IMMUNOASSAY; MEASLES-VIRUS; ANTIBODY; CELLS; PROTEIN; DIAGNOSIS; FUSION; ELISA;
Keywords:
mumps virus; recombinant nucleocapsid protein; electron microscopy; Igm serology;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Clinical Medicine
Life Sciences
Citazioni:
29
Recensione:
Indirizzi per estratti:
Indirizzo: Samuel, D Cent Publ Hlth Lab, Enter Resp & Neurol Virus Lab, 61 Colindale Ave, London NW9 5HT, England Cent Publ Hlth Lab 61 Colindale Ave London England NW9 5HT land
Citazione:
D. Samuel et al., "High level expression of recombinant mumps nucleoprotein in Saccharomyces cerevisiae and its evaluation in mumps IgM serology", J MED VIROL, 66(1), 2002, pp. 123-130

Abstract

To develop improved reagents for mumps serology a high-level yeast expression system was employed to produce recombinant mumps nucleoprotein (rNP). The rNP was purified by CsCl gradient centrifugation and yielded approximately 15 mg/l of yeast culture. Electron microscopy of the rNP revealed characteristic herringbone structures. The electrophoretic mobility of rNP in yeast cells was similar to native NP in SDS-PAGE. Monoclonal antibodies to rNPreacted with native mumps virus nucleoprotein by immunofluorescence assay. A monoclonal antibody to native mumps virus NP reacted with rNP by Westernblot assay. The rNP was investigated as antigen in an IgM capture enzyme immunoassay (EIA) using a horseradish peroxidase conjugate of monoclonal antibody to the rNP. Eighteen sera previously found to be positive by IgM capture radioimmunoassay (MACRIA) and 30 sera that were mumps IgM negative by MACRIA were tested by mumps IgM capture EIA. The results for the two test were concordant. In addition 26 rheumatoid factor positive sera and 35 sera chat were IgM positive for measles, rubella or parvovirus B19 were tested. Fifty-nine sera were negative by mumps IgM capture EIA but two sera collected from two infants 3 and 6 weeks after mumps, measles and rubella vaccination were positive. Mumps MACRIA confirmed these results. Compared to MACRIA the overall sensitivity was 100% (20/20) and specificity was 96.8% (30/31). The yeast expressed rNP was highly immunogenic and suitable for use in IgMcapture EIA for the diagnosis of mumps. (C) 2002 Wiley-Liss, Inc.(dagger)

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 10/07/20 alle ore 18:44:28