Simultaneous Detection of Antibodies to Mouse Hepatitis Virus Recombinant Structural Proteins by a Microsphere-Based Multiplex Fluorescence Immunoassay
| العنوان: | Simultaneous Detection of Antibodies to Mouse Hepatitis Virus Recombinant Structural Proteins by a Microsphere-Based Multiplex Fluorescence Immunoassay |
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| المؤلفون: | Satoshi Kunita, Miyuki Ishida, Tomoko Ishida, Shuko Kameda, Fumihiro Sugiyama, Akira Takakura, Kazuo Goto, Kozue Hagiwara, Ken-ichi Yagami, Kanako Kato |
| المصدر: | Clinical and Vaccine Immunology |
| بيانات النشر: | American Society for Microbiology, 2011. |
| سنة النشر: | 2011 |
| مصطلحات موضوعية: | 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 |
| الوصف: | 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. |
| تدمد: | 1556-679X 1556-6811 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::834c79cd8f18c63b901738cb843be3f9 https://doi.org/10.1128/cvi.00467-10 |
| حقوق: | OPEN |
| رقم الأكسشن: | edsair.doi.dedup.....834c79cd8f18c63b901738cb843be3f9 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 1556679X 15566811 |
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