Development and Large-Scale Testing of a Novel One-Step Triplex RT-qPCR Assay for Simultaneous Detection of “Neurotropic” Porcine Sapeloviruses, Teschoviruses (<i>Picornaviridae</i>) and Type 3 Porcine Astroviruses (<i>Astroviridae</i>) in Various Samples including Nasal Swabs
Porcine sapeloviruses, teschoviruses of family <i>Picornaviridae</i> and type 3 porcine astroviruses of family <i>Astroviridae</i> are (re-)emerging enteric pathogens that could be associated with severe, disseminated infections in swine, affecting multiple organs including t...
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| Format: | Article |
| Language: | English |
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MDPI AG
2022-03-01
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| Series: | Viruses |
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| Online Access: | https://www.mdpi.com/1999-4915/14/3/513 |
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| author | Zoltán László Péter Pankovics Gábor Reuter Attila Cságola Kornélia Bodó Gábor Gáspár Mihály Albert Hunor Bíró Ákos Boros |
| author_facet | Zoltán László Péter Pankovics Gábor Reuter Attila Cságola Kornélia Bodó Gábor Gáspár Mihály Albert Hunor Bíró Ákos Boros |
| author_sort | Zoltán László |
| collection | DOAJ |
| description | Porcine sapeloviruses, teschoviruses of family <i>Picornaviridae</i> and type 3 porcine astroviruses of family <i>Astroviridae</i> are (re-)emerging enteric pathogens that could be associated with severe, disseminated infections in swine, affecting multiple organs including the central nervous system (CNS). Furthermore, small-scale pioneer studies indicate the presence of these viruses in porcine nasal samples to various extents. The laboratory diagnostics are predominantly based on the detection of the viral RNA from faecal and tissue samples using different nucleic-acid-based techniques such as RT-qPCR. In this study, a novel highly sensitive one-step triplex RT-qPCR assay was introduced which can detect all known types of neurotropic sapelo-, tescho- and type 3 astroviruses in multiple types of samples of swine. The assay was evaluated using in vitro synthesized RNA standards and a total of 142 archived RNA samples including known sapelo-, tescho- and type 3 astrovirus positive and negative CNS, enteric and nasal specimens. The results of a large-scale epidemiological investigation of these viruses on <i>n</i> = 473 nasal swab samples from <i>n</i> = 28 industrial-type swine farms in Hungary indicate that all three neurotropic viruses, especially type 3 astroviruses, are widespread and endemically present on most of the investigated farms. |
| first_indexed | 2024-03-09T12:16:40Z |
| format | Article |
| id | doaj.art-44740c619f764f5b864b16252512fce7 |
| institution | Directory Open Access Journal |
| issn | 1999-4915 |
| language | English |
| last_indexed | 2024-03-09T12:16:40Z |
| publishDate | 2022-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Viruses |
| spelling | doaj.art-44740c619f764f5b864b16252512fce72023-11-30T22:45:38ZengMDPI AGViruses1999-49152022-03-0114351310.3390/v14030513Development and Large-Scale Testing of a Novel One-Step Triplex RT-qPCR Assay for Simultaneous Detection of “Neurotropic” Porcine Sapeloviruses, Teschoviruses (<i>Picornaviridae</i>) and Type 3 Porcine Astroviruses (<i>Astroviridae</i>) in Various Samples including Nasal SwabsZoltán László0Péter Pankovics1Gábor Reuter2Attila Cságola3Kornélia Bodó4Gábor Gáspár5Mihály Albert6Hunor Bíró7Ákos Boros8Department of Medical Microbiology and Immunology, Medical School, University of Pécs, 7624 Pécs, HungaryDepartment of Medical Microbiology and Immunology, Medical School, University of Pécs, 7624 Pécs, HungaryDepartment of Medical Microbiology and Immunology, Medical School, University of Pécs, 7624 Pécs, HungaryCeva Phylaxia Ltd., 1107 Budapest, HungaryDepartment of Immunology and Biotechnology, Clinical Center, Medical School, University of Pécs, Szigeti u, 12, 7643 Pécs, HungaryDepartment of Medical Microbiology and Immunology, Medical School, University of Pécs, 7624 Pécs, HungaryCeva Phylaxia Ltd., 1107 Budapest, HungarySHP Ltd., 7400 Kaposvár, HungaryDepartment of Medical Microbiology and Immunology, Medical School, University of Pécs, 7624 Pécs, HungaryPorcine sapeloviruses, teschoviruses of family <i>Picornaviridae</i> and type 3 porcine astroviruses of family <i>Astroviridae</i> are (re-)emerging enteric pathogens that could be associated with severe, disseminated infections in swine, affecting multiple organs including the central nervous system (CNS). Furthermore, small-scale pioneer studies indicate the presence of these viruses in porcine nasal samples to various extents. The laboratory diagnostics are predominantly based on the detection of the viral RNA from faecal and tissue samples using different nucleic-acid-based techniques such as RT-qPCR. In this study, a novel highly sensitive one-step triplex RT-qPCR assay was introduced which can detect all known types of neurotropic sapelo-, tescho- and type 3 astroviruses in multiple types of samples of swine. The assay was evaluated using in vitro synthesized RNA standards and a total of 142 archived RNA samples including known sapelo-, tescho- and type 3 astrovirus positive and negative CNS, enteric and nasal specimens. The results of a large-scale epidemiological investigation of these viruses on <i>n</i> = 473 nasal swab samples from <i>n</i> = 28 industrial-type swine farms in Hungary indicate that all three neurotropic viruses, especially type 3 astroviruses, are widespread and endemically present on most of the investigated farms.https://www.mdpi.com/1999-4915/14/3/513picornavirussapelovirusteschovirusastrovirusmeningoencephalomyelitisswine |
| spellingShingle | Zoltán László Péter Pankovics Gábor Reuter Attila Cságola Kornélia Bodó Gábor Gáspár Mihály Albert Hunor Bíró Ákos Boros Development and Large-Scale Testing of a Novel One-Step Triplex RT-qPCR Assay for Simultaneous Detection of “Neurotropic” Porcine Sapeloviruses, Teschoviruses (<i>Picornaviridae</i>) and Type 3 Porcine Astroviruses (<i>Astroviridae</i>) in Various Samples including Nasal Swabs Viruses picornavirus sapelovirus teschovirus astrovirus meningoencephalomyelitis swine |
| title | Development and Large-Scale Testing of a Novel One-Step Triplex RT-qPCR Assay for Simultaneous Detection of “Neurotropic” Porcine Sapeloviruses, Teschoviruses (<i>Picornaviridae</i>) and Type 3 Porcine Astroviruses (<i>Astroviridae</i>) in Various Samples including Nasal Swabs |
| title_full | Development and Large-Scale Testing of a Novel One-Step Triplex RT-qPCR Assay for Simultaneous Detection of “Neurotropic” Porcine Sapeloviruses, Teschoviruses (<i>Picornaviridae</i>) and Type 3 Porcine Astroviruses (<i>Astroviridae</i>) in Various Samples including Nasal Swabs |
| title_fullStr | Development and Large-Scale Testing of a Novel One-Step Triplex RT-qPCR Assay for Simultaneous Detection of “Neurotropic” Porcine Sapeloviruses, Teschoviruses (<i>Picornaviridae</i>) and Type 3 Porcine Astroviruses (<i>Astroviridae</i>) in Various Samples including Nasal Swabs |
| title_full_unstemmed | Development and Large-Scale Testing of a Novel One-Step Triplex RT-qPCR Assay for Simultaneous Detection of “Neurotropic” Porcine Sapeloviruses, Teschoviruses (<i>Picornaviridae</i>) and Type 3 Porcine Astroviruses (<i>Astroviridae</i>) in Various Samples including Nasal Swabs |
| title_short | Development and Large-Scale Testing of a Novel One-Step Triplex RT-qPCR Assay for Simultaneous Detection of “Neurotropic” Porcine Sapeloviruses, Teschoviruses (<i>Picornaviridae</i>) and Type 3 Porcine Astroviruses (<i>Astroviridae</i>) in Various Samples including Nasal Swabs |
| title_sort | development and large scale testing of a novel one step triplex rt qpcr assay for simultaneous detection of neurotropic porcine sapeloviruses teschoviruses i picornaviridae i and type 3 porcine astroviruses i astroviridae i in various samples including nasal swabs |
| topic | picornavirus sapelovirus teschovirus astrovirus meningoencephalomyelitis swine |
| url | https://www.mdpi.com/1999-4915/14/3/513 |
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