Abstract |
In order to develop sensitive assays for detecting measles antibodies in oral fluid specimens, we have produced recombinant measles virus nucleoprotein (rMVN) in a yeast expression system and prepared monoclonal antibodies to the protein. Measles nucleoprotein gene from the Schwarz vaccine strain was cloned into a yeast expression vector, pFX7 under the control of the hybrid GAL10-PYK1 promoter. High levels of rMVN (20 mg/litre of yeast culture) were generated. Electron microscopy showed that the purified rMVN assembled into typical herring-bone structures. Monoclonal antibodies produced to the rMVN also reacted with native measles virus N in immunofluorescence tests. The purified rMVN and a monoclonal antibody to the rMVN conjugated to horseradish peroxidase were used to develop a measles specific IgM capture EIA (MACEIA) in both serum and oral fluid specimens. Evaluations of the MACEIA were performed by testing a) serum samples (n=80) and b) paired oral fluid/serum samples from measles cases (n=50, representing 16 cases) and oral fluids from controls with non- measles rash (n=59, representing 48 cases). The samples were also tested for measles IgM, using a reference radioimmunoassay (MACRIA). The sensitivity and specificity of the MACEIA compared with MACRIA for a) the serum samples were 100 and 96.6% respectively and b) for paired serum/oral fluids samples 100 and 100%, respectively.
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Authors | Dhanraj Samuel, Kestutis Sasnauskas, Li Jin, Alma Gedvilaite, Rimas Slibinskas, Stuart Beard, Aurelija Zvirbliene, Solange Artimos Oliveira, Juozas Staniulis, Bernard Cohen, David Brown |
Journal | Journal of clinical virology : the official publication of the Pan American Society for Clinical Virology
(J Clin Virol)
Vol. 28
Issue 2
Pg. 121-9
(Oct 2003)
ISSN: 1386-6532 [Print] Netherlands |
PMID | 12957182
(Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
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Chemical References |
- Antibodies, Viral
- Immunoglobulin M
- Nucleoproteins
- Recombinant Proteins
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Topics |
- Animals
- Antibodies, Viral
- Enzyme-Linked Immunosorbent Assay
(methods)
- Humans
- Immunoglobulin M
- Measles
(diagnosis)
- Measles virus
(immunology)
- Mice
- Mice, Inbred BALB C
- Nucleoproteins
(genetics, immunology, metabolism)
- Recombinant Proteins
(genetics, immunology, metabolism)
- Saccharomyces cerevisiae
(genetics, metabolism)
- Saliva
(immunology)
- Sensitivity and Specificity
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