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Simultaneous Detection of Antibodies to Mouse Hepatitis Virus Recombinant Structural Proteins by a Microsphere-Based Multiplex Fluorescence Immunoassay
- Source :
- Clinical and Vaccine Immunology
- Publication Year :
- 2011
- Publisher :
- American Society for Microbiology, 2011.
-
Abstract
- We describe a new microsphere-based multiplex fluorescent immunoassay (MFI) using recombinant mouse hepatitis virus (MHV) proteins to detect antibodies to coronaviruses in mouse and rat sera. All the recombinant proteins, including nucleocapsid (N) and 3 subunits of spike protein, S1, S2, and Smid, showed positive reactivity in MFI with mouse antisera to 4 MHV strains (MHV-S, -A59, -JHM, and -Nu67) and rat antiserum to a strain of sialodacryoadenitis virus (SDAV-681). The MFI was evaluated for its diagnostic power, with panels of mouse sera classified as positive or negative for anti-MHV antibodies by enzyme-linked immunosorbent assay (ELISA) using MHV virion antigen and indirect fluorescent antibody assay. The reactivities of 236 naturally infected mouse sera were examined; 227 samples were positive by MFI using S2 antigen (96% sensitivity), and 208 samples were positive using N antigen (88% sensitivity). Based on the assessment by MFI using the S2 and N antigens, only 3 serum samples showed double-negative results, indicating a false-negative rate of 1.3%. In 126 uninfected mouse sera, including 34 ELISA false-positive sera, only 7 samples showed false-positive results by MFI using either the S2 or N antigen (94% specificity). Similarly, the S2 and N antigen-based MFI was 98% sensitive and 100% specific in detecting anticoronavirus antibodies in rat sera. Thus, this MFI-based serologic assay using the S2 and N antigens promises to be a reliable diagnostic method, representing a highly sensitive and specific alternative to traditional ELISA for detection of coronavirus infections in laboratory mouse and rat colonies.
- Subjects :
- Microbiology (medical)
viruses
Molecular Sequence Data
Clinical Biochemistry
Immunology
Antibodies, Viral
Sensitivity and Specificity
Veterinary Immunology
Fluorescence
law.invention
Serology
Rodent Diseases
Mice
Mouse hepatitis virus
Antigen
law
medicine
Animals
Immunology and Allergy
Multiplex
Immunoassay
Antiserum
Murine hepatitis virus
biology
medicine.diagnostic_test
Clinical Laboratory Techniques
Sequence Analysis, DNA
biology.organism_classification
Virology
Molecular biology
Microspheres
Rats
Recombinant DNA
biology.protein
RNA, Viral
Female
Antibody
Coronavirus Infections
Subjects
Details
- ISSN :
- 1556679X and 15566811
- Volume :
- 18
- Database :
- OpenAIRE
- Journal :
- Clinical and Vaccine Immunology
- Accession number :
- edsair.doi.dedup.....834c79cd8f18c63b901738cb843be3f9