4
views
0
recommends
+1 Recommend
0 collections
    0
    shares
      • Record: found
      • Abstract: found
      • Article: not found

      Broad respiratory virus detection in infants hospitalized for bronchiolitis by use of a multiplex RT‐PCR DNA microarray system†

      research-article

      Read this article at

      ScienceOpenPublisherPMC
      Bookmark
          There is no author summary for this article yet. Authors can add summaries to their articles on ScienceOpen to make them more accessible to a non-specialist audience.

          Abstract

          Newly available molecular tools allow a sensitive detection of a broad panel of viruses in respiratory tract specimens. In the present study, the application of a multiplex RT‐PCR DNA microarray in diagnosis and epidemiological survey of viral infections in infants hospitalized for bronchiolitis was assessed. One hundred and thirty‐eight nasopharyngeal aspirates collected from October 2007 to September 2008 were tested by direct immunofluorescence and viral culture, a combination of referenced RT‐PCRs and the DNA microarray. One or more viruses were detected in 96, 126 and 126 of the specimens by direct immunofluorescence and viral culture, RT‐PCRs and DNA microarray, respectively (70 vs. 91 vs. 91%, P < 10 −3). The RT‐PCRs and the DNA microarray yielded concordant results for 99% of specimens and identified mixed viral infections in 85 (62%). The most common associations were: human bocavirus and respiratory syncytial virus (32%), adenovirus and respiratory syncytial virus (30%), and parainfluenza virus type 3 and respiratory syncytial virus (23%). None of the bronchiolitis severity parameters including intensive care unit admission, O 2 supply, O 2 saturation percentage, O 2 length and length of stay at the hospital appeared to be significantly increased in multiple viral infections compared to single viral infections ( P > 0.1). In conclusion, the use of this DNA microarray in clinical virology practice allows rapid and accurate identification of common and uncommon viral respiratory pathogens in infants hospitalized for bronchiolitis. It should improve the clinical management, the epidemiological survey, and the prevention of the nosocomial transmission of respiratory viruses in pediatric wards. J. Med. Virol. 84:979–985, 2012. © 2012 Wiley Periodicals, Inc.

          Related collections

          Most cited references29

          • Record: found
          • Abstract: found
          • Article: not found

          Detection of respiratory viruses by molecular methods.

          Clinical laboratories historically diagnose seven or eight respiratory virus infections using a combination of techniques including enzyme immunoassay, direct fluorescent antibody staining, cell culture, and nucleic acid amplification tests. With the discovery of six new respiratory viruses since 2000, laboratories are faced with the challenge of detecting up to 19 different viruses that cause acute respiratory disease of both the upper and lower respiratory tracts. The application of nucleic acid amplification technology, particularly multiplex PCR coupled with fluidic or fixed microarrays, provides an important new approach for the detection of multiple respiratory viruses in a single test. These multiplex amplification tests provide a sensitive and comprehensive approach for the diagnosis of respiratory tract infections in individual hospitalized patients and the identification of the etiological agent in outbreaks of respiratory tract infection in the community. This review describes the molecular methods used to detect respiratory viruses and discusses the contribution that molecular testing, especially multiplex PCR, has made to our ability to detect respiratory viruses and to increase our understanding of the roles of various viral agents in acute respiratory disease.
            Bookmark
            • Record: found
            • Abstract: found
            • Article: not found

            Dual Infection of Infants by Human Metapneumovirus and Human Respiratory Syncytial Virus Is Strongly Associated with Severe Bronchiolitis

            Abstract The association between severe bronchiolitis and dual infection by human metapneumovirus (hMPV) and human respiratory syncytial virus (hRSV) was investigated in !2-year-old infants with bronchiolitis who were admitted to the hospital during the 2001–2002 winter season. hMPV in nasopharyngeal aspirate and/or cells and fluid collected by nonbronchoscopic bronchoalveolar lavage was detected by reverse transcriptase-polymerase chain reaction (RT-PCR). hRSV was detected in nasopharyngeal aspirate and/or cells and fluid collected by nonbronchoscopic bronchoalveolar lavage by enzyme immunoassay, tissue culture, and RT-PCR. Dual infection with hMPV and hRSV confers a 10-fold increase in relative risk (RR) of admission to a pediatric intensive-care unit for mechanical ventilation (RR, 10.99 [95% confidence interval, 5.0–24.12]; P < .001, by Fisher exact test). Dual infection by hMPV and hRSV is associated with severe bronchiolitis.
              Bookmark
              • Record: found
              • Abstract: found
              • Article: not found

              Simultaneous detection of fourteen respiratory viruses in clinical specimens by two multiplex reverse transcription nested‐PCR assays

              Abstract There is a need for rapid, sensitive, and accurate diagnosis of lower respiratory tract infections in children, elderly, and immunocompromised patients, who are susceptible to serious complications. The multiplex RT‐nested PCR assay has been used widely for simultaneous detection of non‐related viruses involved in infectious diseases because of its high specificity and sensitivity. A new multiplex RT‐PCR assay is described in this report. This approach includes nested primer sets targeted to conserve regions of human parainfluenza virus haemagglutinin, human coronavirus spike protein, and human enterovirus and rhinovirus polyprotein genes. It permits rapid, sensitive, and simultaneous detection and typing of the four types of parainfluenza viruses (1, 2, 3, 4AB), human coronavirus 229E and OC43, and the generic detection of enteroviruses and rhinoviruses. The testing of 201 clinical specimens with this multiplex assay along with other one formerly described by our group to simultaneously detect and type the influenza viruses, respiratory syncytial viruses, and a generic detection of all serotypes of adenovirus, covers the detection of most viruses causing respiratory infectious disease in humans. The results obtained were compared with conventional viral culture, immunofluorescence assay, and a third multiplex RT‐PCR assay for all human parainfluenza viruses types described previously. In conclusion, both multiplex RT‐PCR assays provide a system capable of detecting and identifying simultaneously 14 different respiratory viruses in clinical specimens with high sensitivity and specificity, being useful for routine diagnosis and survey of these viruses within the population. J. Med. Virol. 72:484–495, 2004. © 2004 Wiley‐Liss, Inc.
                Bookmark

                Author and article information

                Contributors
                landreoletti@chu-reims.fr
                Journal
                J Med Virol
                J. Med. Virol
                10.1002/(ISSN)1096-9071
                JMV
                Journal of Medical Virology
                Wiley Subscription Services, Inc., A Wiley Company (Hoboken )
                0146-6615
                1096-9071
                June 2012
                12 April 2012
                : 84
                : 6 ( doiID: 10.1002/jmv.v84.6 )
                : 979-985
                Affiliations
                [ 1 ]Medical and Molecular Virology Unit and EA‐484, University Hospital Center and Medical School of Reims, Reims, France
                [ 2 ]Pediatric Department, American Memorial Hospital, University Hospital Center of Reims, Reims, France
                [ 3 ]UMR‐S 707 Medical School of St. Antoine, Paris, France
                Author notes
                [*] [* ]Laboratoire de Virologie Médicale et Moléculaire, et EA, Hôpital Robert Debré, Avenue du Général Koenig, 51092 REIMS Cedex, France.===
                Article
                JMV23272
                10.1002/jmv.23272
                7166763
                22499022
                43140fe5-7ed0-440a-af22-99f7eb08ecc8
                Copyright © 2012 Wiley Periodicals, Inc.

                This article is being made freely available through PubMed Central as part of the COVID-19 public health emergency response. It can be used for unrestricted research re-use and analysis in any form or by any means with acknowledgement of the original source, for the duration of the public health emergency.

                History
                : 15 February 2012
                Page count
                Figures: 1, Tables: 3, References: 31, Pages: 6, Words: 6328
                Funding
                Funded by: Reims University Medical Centre
                Funded by: French Army Department (Bourse DGA: Délégation Générale de l'Armement, Ministère de la Défense, Topic: Microbiology, infectious diseases; to F.R.)
                Categories
                Research Article
                Research Articles
                Custom metadata
                2.0
                June 2012
                Converter:WILEY_ML3GV2_TO_JATSPMC version:5.8.0 mode:remove_FC converted:15.04.2020

                Microbiology & Virology
                bronchiolitis,infants,rt‐pcr dna microarray,respiratory viruses,clinical severity parameters

                Comments

                Comment on this article