<span style="font-variant: small-caps">“FastCheck</span><sup>FLI</sup> PPR-like”—A Molecular Tool for the Fast Genome Detection of PPRV and Differential Diagnostic Pathogens
To assist the global eradication of peste des petits ruminants virus (PPRV), a molecular test for the rapid and reliable detection of PPRV was developed which additionally enables the detection of pathogens relevant for differential diagnostics. For this purpose, the necessary time frame of a magnet...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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MDPI AG
2020-10-01
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| Series: | Viruses |
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| Online Access: | https://www.mdpi.com/1999-4915/12/11/1227 |
| _version_ | 1797549358169718784 |
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| author | Sabrina Halecker Thomas C. Mettenleiter Martin Beer Bernd Hoffmann |
| author_facet | Sabrina Halecker Thomas C. Mettenleiter Martin Beer Bernd Hoffmann |
| author_sort | Sabrina Halecker |
| collection | DOAJ |
| description | To assist the global eradication of peste des petits ruminants virus (PPRV), a molecular test for the rapid and reliable detection of PPRV was developed which additionally enables the detection of pathogens relevant for differential diagnostics. For this purpose, the necessary time frame of a magnetic bead-based nucleic acid extraction protocol was markedly shortened to 7 min and 13 s. The optimized extraction was run on a BioSprint 15 platform. Furthermore, a high-speed multi-well RT-qPCR for the genome detection of PPRV and additional important pathogens such as Foot-and-mouth disease virus, Parapoxvirus ovis, Goatpox virus, and <i>Mycoplasma capricolum subsp. capripneumoniae</i> was established and combined with suitable internal control assays. The here-described qPCR is based on a lyophilized master mix and takes only around 30 to 40 min. Several qPCR cyclers were evaluated regarding their suitability for fast-cycling approaches and for their diagnostic performance in a high-speed RT-qPCR. The final evaluation was conducted on the BioRad CFX96 and also on a portable Liberty16 qPCR cycler. The new molecular test designated as “<span style="font-variant: small-caps;">FastCheck</span><sup>FLI</sup> PPR-like”, which is based on rapid nucleic acid extraction and high-speed RT-qPCR, delivered reliable results in less than one hour, allowing its use also in a pen-side scenario. |
| first_indexed | 2024-03-10T15:13:34Z |
| format | Article |
| id | doaj.art-11341eea0b544f00916439b1b2dfdb0a |
| institution | Directory Open Access Journal |
| issn | 1999-4915 |
| language | English |
| last_indexed | 2024-03-10T15:13:34Z |
| publishDate | 2020-10-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Viruses |
| spelling | doaj.art-11341eea0b544f00916439b1b2dfdb0a2023-11-20T19:05:19ZengMDPI AGViruses1999-49152020-10-011211122710.3390/v12111227<span style="font-variant: small-caps">“FastCheck</span><sup>FLI</sup> PPR-like”—A Molecular Tool for the Fast Genome Detection of PPRV and Differential Diagnostic PathogensSabrina Halecker0Thomas C. Mettenleiter1Martin Beer2Bernd Hoffmann3Institute of Diagnostic Virology, Friedrich-Loeffler-Institut, Südufer 10, 17493 Greifswald-Insel Riems, GermanyFriedrich-Loeffler-Institut, Südufer 10, 17493 Greifswald-Insel Riems, GermanyInstitute of Diagnostic Virology, Friedrich-Loeffler-Institut, Südufer 10, 17493 Greifswald-Insel Riems, GermanyInstitute of Diagnostic Virology, Friedrich-Loeffler-Institut, Südufer 10, 17493 Greifswald-Insel Riems, GermanyTo assist the global eradication of peste des petits ruminants virus (PPRV), a molecular test for the rapid and reliable detection of PPRV was developed which additionally enables the detection of pathogens relevant for differential diagnostics. For this purpose, the necessary time frame of a magnetic bead-based nucleic acid extraction protocol was markedly shortened to 7 min and 13 s. The optimized extraction was run on a BioSprint 15 platform. Furthermore, a high-speed multi-well RT-qPCR for the genome detection of PPRV and additional important pathogens such as Foot-and-mouth disease virus, Parapoxvirus ovis, Goatpox virus, and <i>Mycoplasma capricolum subsp. capripneumoniae</i> was established and combined with suitable internal control assays. The here-described qPCR is based on a lyophilized master mix and takes only around 30 to 40 min. Several qPCR cyclers were evaluated regarding their suitability for fast-cycling approaches and for their diagnostic performance in a high-speed RT-qPCR. The final evaluation was conducted on the BioRad CFX96 and also on a portable Liberty16 qPCR cycler. The new molecular test designated as “<span style="font-variant: small-caps;">FastCheck</span><sup>FLI</sup> PPR-like”, which is based on rapid nucleic acid extraction and high-speed RT-qPCR, delivered reliable results in less than one hour, allowing its use also in a pen-side scenario.https://www.mdpi.com/1999-4915/12/11/1227peste des petits ruminants virus (PPRV)<i>Small ruminant morbilli virus</i>molecular pen-side testfast extractionhigh-speed RT-qPCRrapid detection method |
| spellingShingle | Sabrina Halecker Thomas C. Mettenleiter Martin Beer Bernd Hoffmann <span style="font-variant: small-caps">“FastCheck</span><sup>FLI</sup> PPR-like”—A Molecular Tool for the Fast Genome Detection of PPRV and Differential Diagnostic Pathogens Viruses peste des petits ruminants virus (PPRV) <i>Small ruminant morbilli virus</i> molecular pen-side test fast extraction high-speed RT-qPCR rapid detection method |
| title | <span style="font-variant: small-caps">“FastCheck</span><sup>FLI</sup> PPR-like”—A Molecular Tool for the Fast Genome Detection of PPRV and Differential Diagnostic Pathogens |
| title_full | <span style="font-variant: small-caps">“FastCheck</span><sup>FLI</sup> PPR-like”—A Molecular Tool for the Fast Genome Detection of PPRV and Differential Diagnostic Pathogens |
| title_fullStr | <span style="font-variant: small-caps">“FastCheck</span><sup>FLI</sup> PPR-like”—A Molecular Tool for the Fast Genome Detection of PPRV and Differential Diagnostic Pathogens |
| title_full_unstemmed | <span style="font-variant: small-caps">“FastCheck</span><sup>FLI</sup> PPR-like”—A Molecular Tool for the Fast Genome Detection of PPRV and Differential Diagnostic Pathogens |
| title_short | <span style="font-variant: small-caps">“FastCheck</span><sup>FLI</sup> PPR-like”—A Molecular Tool for the Fast Genome Detection of PPRV and Differential Diagnostic Pathogens |
| title_sort | span style font variant small caps fastcheck span sup fli sup ppr like a molecular tool for the fast genome detection of pprv and differential diagnostic pathogens |
| topic | peste des petits ruminants virus (PPRV) <i>Small ruminant morbilli virus</i> molecular pen-side test fast extraction high-speed RT-qPCR rapid detection method |
| url | https://www.mdpi.com/1999-4915/12/11/1227 |
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