期刊
JOURNAL OF VIROLOGICAL METHODS
卷 254, 期 -, 页码 51-64出版社
ELSEVIER
DOI: 10.1016/j.jviromet.2018.01.011
关键词
Monoclonal antibodies; Clinical samples; Infected cells; ELISA; hRSV; hMPV; ADV; Immunofluorescense; Dot blot; Purified proteins
资金
- National Fund for Scientific and Technological Development (FONDECYT) program of the Ministry of Education of Chile [1140010, 1150862, 1131012]
- INNOVA-CORFO program of Chilean Ministry of Economy [11IDL2-10668, 15VEIID-45704, 13CTI-21526 P5]
- Millennium Institute in Immunology and Immunotherapy of the Ministry of Economy of Chile [P09-016-F]
- Pediatric Division and School of Medicine, Pontificia Universidad Catolica de Chile
Human Respiratory Syncytial Virus (hRSV), human Metapneumovirus (hMPV) and Adenovirus (ADV), are three of the most prevalent viruses responsible for pneumonia and bronchiolitis in children and elderly worldwide, accounting for a high number of hospitalizations annually. Diagnosis of these viruses is required to take clinical actions that allow an appropriate patient management. Thereby, new strategies to design fast diagnostic methods are highly required. In the present work, six monoclonal antibodies (mAbs, two for each virus) specific for conserved proteins from hRSV, hMPV and ADV were generated and evaluated through different immunological techniques, based on detection of purified protein, viral particles and human samples. In vitro evaluation of these antibodies showed higher specificity and sensitivity than commercial antibodies tested in this study. These antibodies were used to design a sandwich ELISA tests that allowed the detection of hRSV, hMPV, and ADV in human nasopharyngeal swabs. We observed that hRSV and ADV were detected with sensitivity and specificity equivalent to a current Direct Fluorescence Assay (DFA) methodology. However, hMPV was detected with more sensitivity than DFA. Our data suggest that these new mAbs can efficiently identify infected samples and discriminate from patients infected with other respiratory pathogens.
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