Rapid and sensitive detection of metapneumovirus in clinical specimens by indirect fluorescence assay using a monoclonal antibody

• Published in: Journal of medical virology 2008, Vol.80 (1), p.154-158
• Main Authors: Manoha, C, Bour, J.B, Pitoiset, C, Darniot, M, Aho, S, Pothier, P
• Format: Article
• Language: English
• Subjects: Adult; Aged; Animals; Antibodies, Monoclonal; Antibodies, Monoclonal - immunology; Antigens, Viral; Antigens, Viral - analysis; Biological and medical sciences; Cells, Cultured; Child; diagnostic; Fluorescent Antibody Technique, Indirect; Fluorescent Antibody Technique, Indirect - methods; Fundamental and applied biological sciences. Psychology; genotype; Human metapneumovirus; Human viral diseases; Humans; immunofluorescence; Infectious diseases; Influenza virus; Life Sciences; Medical sciences; Metapneumovirus; Metapneumovirus - immunology; Metapneumovirus - isolation & purification; Mice; Mice, Inbred BALB C; Microbiology; Microbiology and Parasitology; Miscellaneous; monoclonal antibody; Paramyxoviridae Infections; Paramyxoviridae Infections - diagnosis; Respiratory syncytial virus; Respiratory Tract Infections; Respiratory Tract Infections - diagnosis; Respiratory Tract Infections - immunology; Sensitivity and Specificity; Vaccines, antisera, therapeutical immunoglobulins and monoclonal antibodies; Viral diseases; Virology
• ISSN: 0146-6615; 1096-9071
• DOI: 10.1002/jmv.21038

Human metapneumovirus, with two known genotypes named A and B, is associated with mild respiratory symptoms to severe LRTI in children, high-risk adults and the elderly. Rapid and reliable methods of hMPV detection in clinical samples are essential to implement appropriate care, to better understand the pathology of hMPV and to determine its epidemiology. Respiratory samples from 1,386 patients collected during 2 consecutive years were screened for hMPV using indirect immunofluorescence (IFA) assay with a monoclonal antibody. Forty-three patients tested positive for hMPV by the IFA method. In parallel, the samples were examined with RT-PCR on the F gene. Of these, 41 specimens were RT-PCR positive. The remaining two IF positives were cultured and the cultures were subsequently RT-PCR positive. IFA showed therefore a sensitivity of 100%. No false positive signals were obtained with the influenza virus, respiratory syncytial virus or parainfluenza. When tested by RT-PCR, all IFA-negative samples (n = 204)were found negative. Therefore the specificity of IFA was 100%, IC₉₅ [98-100%], with a negative predictive value of 100%. Based upon phylogenetic analysis of the fusion gene, both subgroups of hMPV were efficiently detected by IFA, and the viral aetiology could be given in 2 hr. These results demonstrate the potential usefulness of immunofluorescence with our monoclonal antibody for the rapid detection of hMPV in clinical specimens in the management of therapy and the control of nosocomial diffusion. J. Med. Virol. 80:154-158, 2008. © 2007 Wiley-Liss, Inc.