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      Development of a Rapid Fluorescent Immunochromatographic Test to Detect Respiratory Syncytial Virus

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          Abstract

          Human respiratory syncytial virus (RSV) is one of the most common viruses infecting the respiratory tracts of infants. The rapid and sensitive detection of RSV is important to minimize the incidence of infection. In this study, novel monoclonal antibodies (mAbs; B11A5 and E8A11) against RSV nucleoprotein (NP) were developed and applied to develop a rapid fluorescent immunochromatographic strip test (FICT), employing europium nanoparticles as the fluorescent material. For the FICT, the limits of detection of the antigen and virus were 1.25 µg/mL and 4.23 × 10 6 TCID 50/mL, respectively, corresponding to 4.75 × 10 6 ± 5.8 ×10 5 (mean ± SD) RNA copy numbers per reaction mixture for RSV NP. A clinical study revealed a sensitivity of 90% (18/20) and specificity of 98.18% (108/110) for RSV detection when comparing the performance to that of reverse transcription polymerase chain reaction (RT-PCR), representing a 15% improvement in sensitivity over the SD Bioline rapid kit. This newly developed FICT could be a useful tool for the rapid diagnosis of RSV infection.

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          Limit of blank, limit of detection and limit of quantitation.

          David Armbruster, Terry Pry (2008)
          * Limit of Blank (LoB), Limit of Detection (LoD), and Limit of Quantitation (LoQ) are terms used to describe the smallest concentration of a measurand that can be reliably measured by an analytical procedure. * LoB is the highest apparent analyte concentration expected to be found when replicates of a blank sample containing no analyte are tested. LoB = mean(blank) + 1.645(SD(blank)). * LoD is the lowest analyte concentration likely to be reliably distinguished from the LoB and at which detection is feasible. LoD is determined by utilising both the measured LoB and test replicates of a sample known to contain a low concentration of analyte. * LoD = LoB + 1.645(SD (low concentration sample)). * LoQ is the lowest concentration at which the analyte can not only be reliably detected but at which some predefined goals for bias and imprecision are met. The LoQ may be equivalent to the LoD or it could be at a much higher concentration.
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            ICTV Virus Taxonomy Profile: Pneumoviridae.

            Benhur Lee, Linfa Wang, Ictv Report Consortium … (2017)
            The family Pneumoviridae comprises large enveloped negative-sense RNA viruses. This taxon was formerly a subfamily within the Paramyxoviridae, but was reclassified in 2016 as a family with two genera, Orthopneumovirus and Metapneumovirus. Pneumoviruses infect a range of mammalian species, while some members of the Metapneumovirus genus may also infect birds. Some viruses are specific and pathogenic for humans, such as human respiratory syncytial virus and human metapneumovirus. There are no known vectors for pneumoviruses and transmission is thought to be primarily by aerosol droplets and contact. This is a summary of the International Committee on Taxonomy of Viruses (ICTV) Report on the taxonomy of the Pneumoviridae, which is available at www.ictv.global/report/pneumoviridae.
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              Cross-neutralization of four paramyxoviruses by a human monoclonal antibody.

              B Guarino, Benjamin J Marsland, Federica Sallusto … (2013)
              Broadly neutralizing antibodies reactive against most and even all variants of the same viral species have been described for influenza and HIV-1 (ref. 1). However, whether a neutralizing antibody could have the breadth of range to target different viral species was unknown. Human respiratory syncytial virus (HRSV) and human metapneumovirus (HMPV) are common pathogens that cause severe disease in premature newborns, hospitalized children and immune-compromised patients, and play a role in asthma exacerbations. Although antisera generated against either HRSV or HMPV are not cross-neutralizing, we speculated that, because of the repeated exposure to these viruses, cross-neutralizing antibodies may be selected in some individuals. Here we describe a human monoclonal antibody (MPE8) that potently cross-neutralizes HRSV and HMPV as well as two animal paramyxoviruses: bovine RSV (BRSV) and pneumonia virus of mice (PVM). In its germline configuration, MPE8 is HRSV-specific and its breadth is achieved by somatic mutations in the light chain variable region. MPE8 did not result in the selection of viral escape mutants that evaded antibody targeting and showed potent prophylactic efficacy in animal models of HRSV and HMPV infection, as well as prophylactic and therapeutic efficacy in the more relevant model of lethal PVM infection. The core epitope of MPE8 was mapped on two highly conserved anti-parallel β-strands on the pre-fusion viral F protein, which are rearranged in the post-fusion F protein conformation. Twenty-six out of the thirty HRSV-specific neutralizing antibodies isolated were also found to be specific for the pre-fusion F protein. Taken together, these results indicate that MPE8 might be used for the prophylaxis and therapy of severe HRSV and HMPV infections and identify the pre-fusion F protein as a candidate HRSV vaccine.
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                Author and article information

                Journal
                Journal ID (nlm-ta): Int J Mol Sci
                Journal ID (iso-abbrev): Int J Mol Sci
                Journal ID (publisher-id): ijms
                Title: International Journal of Molecular Sciences
                Publisher: MDPI
                ISSN (Electronic): 1422-0067
                Publication date (Electronic): 02 October 2018
                Publication date Collection: October 2018
                Volume: 19
                Issue: 10
                Electronic Location Identifier: 3013
                Affiliations
                [1 ]Zoonosis Research Center, Department of Infection Biology, School of Medicine, Wonkwang University, 460, Iksan-daero, Iksan 54538, Korea; trinhthithuytien.k56@ 123456hus.edu.vn (T.T.T.T.); hyunpk@ 123456wku.ac.kr (H.P.); tuonghien23@ 123456gmail.com (H.T.T.)
                [2 ]Department of Pediatrics, School of Medicine, Wonkwang University, 460, Iksan-daero, Iksan 54538, Korea; yudoc@ 123456wku.ac.kr (S.-T.Y.); cdy8118@ 123456wku.ac.kr (D.-Y.C.)
                Author notes
                [* ]Correspondence: yeosj@ 123456wku.ac.kr ; Tel.: +82-63-850-6777; Fax: +82-63-857-0342
                [†]

                These authors contributed equally to this manuscript.

                Article
                Publisher ID: ijms-19-03013
                DOI: 10.3390/ijms19103013
                PMC ID: 6212954
                PubMed ID: 30279406
                SO-VID: 523f97b7-8912-496b-98fd-c7637d4eb407
                Copyright © © 2018 by the authors.
                License:

                Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license ( http://creativecommons.org/licenses/by/4.0/).

                History
                Date received : 19 August 2018
                Date accepted : 29 September 2018
                Categories
                Subject: Article

                ScienceOpen disciplines: Molecular biology
                Keywords: monoclonal antibody,respiratory syncytial virus,nucleoprotein,fluorescent immunochromatographic test,clinical study
                Data availability:
                ScienceOpen disciplines: Molecular biology
                Keywords: monoclonal antibody, respiratory syncytial virus, nucleoprotein, fluorescent immunochromatographic test, clinical study

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