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      Multiplex detection in tonsillar tissue of all known human polyomaviruses

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          Abstract

          Background

          In the past few years, eleven new human viruses have joined the two previously known members JCPyV and BKPyV of the Polyomaviridae family, by virtue of molecular methods. Serology data suggest that infections with human polyomaviruses (HPyVs) occur since childhood and the viruses are widespread in the general population. However, the viral persistence sites and transmission routes are by and large unknown. Our previous studies demonstrated that the four new HPyVs – KIPyV, WUPyV, MCPyV and TSPyV – were present in the tonsils, and suggested lymphoid tissue as a persistent site of these emerging human viruses.

          We developed a Luminex-based multiplex assay for simultaneous detection of all 13 HPyVs known, and explored their occurrence in tonsillar tissues of children and adults mostly with tonsillitis or tonsillar hypertrophy.

          Methods

          We set up and validated a new Luminex-based multiplex assay by using primer pairs and probes targeting the respective HPyV viral protein 1 (VP1) genes. With this assay we tested 78 tonsillar tissues for DNAs of 13 HPyVs.

          Results

          The multiplex assay allowed for simultaneous detection of 13 HPyVs with high analytical sensitivity and specificity, with detection limits of 10 0–10 2 copies per microliter, and identified correctly all 13 target sequences with no cross reactions. HPyV DNA altogether was found in 14 (17.9%) of 78 tonsils. The most prevalent HPyVs were HPyV6 (7.7%), TSPyV (3.8%) and WUPyV (3.8%). Mixed infection of two HPyVs occurred in one sample.

          Conclusions

          The Luminex-based HPyV multiplex assay appears highly suitable for clinical diagnostic purposes and large-scale epidemiological studies. Additional evidence was acquired that the lymphoid system plays a role in HPyV infection and persistence. Thereby, shedding from this site during reactivation might take part in transmission of the newly found HPyVs.

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          Most cited references16

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          New human papovavirus (B.K.) isolated from urine after renal transplantation.

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            Cultivation of papova-like virus from human brain with progressive multifocal leucoencephalopathy.

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              Seroepidemiology of the human polyomaviruses.

              To assess the stability of polyomavirus antibodies in serial samples over time and the incidence and age-specific prevalence of polyomavirus infections, we established enzyme immunoassays (EIAs) using purified yeast-expressed virus-like particles (VLPs) containing the VP1 major capsid proteins of JC virus (JCV) and the AS and SB strains of BK virus (BKV). A random subsample of 150 Finnish women who had serum samples taken during the first trimester of pregnancy and had a second pregnancy during a 5 year follow-up period was selected, grouped by age of first pregnancy. The polyomavirus antibody levels were similar in samples taken during the first and second pregnancies (correlation coefficient 0.93 for BKV SB and 0.94 for JCV). Analysis of serum samples from 290 Swedish children aged 1-13 years, grouped by age in 2 year intervals, demonstrated that BKV seropositivity increased rapidly with increasing age of the children, reaching 98 % seroprevalence at 7-9 years of age, followed by a minor decrease. JCV seroprevalence increased only slowly with increasing age and reaching 72 % positivity among mothers >25 years of age. The age-specific seroprevalence of the human polyomaviruses measured using this VLP-based EIA was similar to previous serosurveys by other methods. The stability of the antibodies over time indicates that polyomavirus seropositivity is a valid marker of cumulative virus exposure, and polyoma VLP-based EIAs may therefore be useful for epidemiological studies of these viruses.
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                Author and article information

                Contributors
                rsadeghi@bloodsystems.org
                yilin.wang@helsinki.fi
                torbjorn.ramqvist@ki.se
                leena-maija.aaltonen@hus.fi
                lari.pyoria@helsinki.fi
                mari.toppinen@helsinki.fi
                maria.soderlund-venermo@helsinki.fi
                +358-5052-49086 , klaus.hedman@helsinki.fi
                Journal
                BMC Infect Dis
                BMC Infect. Dis
                BMC Infectious Diseases
                BioMed Central (London )
                1471-2334
                8 June 2017
                8 June 2017
                2017
                : 17
                : 409
                Affiliations
                [1 ]ISNI 0000 0004 0410 2071, GRID grid.7737.4, Virology, , University of Helsinki, ; Helsinki, Finland
                [2 ]ISNI 0000 0004 1937 0626, GRID grid.4714.6, Department of Oncology-Pathology, , Karolinska Institutet, ; Stockholm, Sweden
                [3 ]ISNI 0000 0000 9950 5666, GRID grid.15485.3d, Department of Otorhinolaryngology-Head and Neck Surgery, , Helsinki University Hospital, ; Helsinki, Finland
                [4 ]ISNI 0000 0004 0410 2071, GRID grid.7737.4, , University of Helsinki, ; Helsinki, Finland
                [5 ]ISNI 0000 0000 9950 5666, GRID grid.15485.3d, , Helsinki University Hospital, HUSLAB, ; Helsinki, Finland
                Author information
                http://orcid.org/0000-0002-6947-3012
                Article
                2479
                10.1186/s12879-017-2479-5
                5465560
                28595595
                b20ca851-d148-488b-b01c-eac2b6b70962
                © The Author(s). 2017

                Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License ( http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver ( http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

                History
                : 23 November 2016
                : 22 May 2017
                Categories
                Research Article
                Custom metadata
                © The Author(s) 2017

                Infectious disease & Microbiology
                hpyv,pcr,luminex,tonsil
                Infectious disease & Microbiology
                hpyv, pcr, luminex, tonsil

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