Development of a recombinase polymerase amplification assay for viral haemorrhagic septicemia virus
© 2022 John Wiley & Sons Ltd.
| Veröffentlicht in: | Journal of fish diseases. - 1998. - 45(2022), 8 vom: 25. Aug., Seite 1065-1071 |
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| Weitere Verfasser: | , , , |
| Format: | Online-Aufsatz |
| Sprache: | English |
| Veröffentlicht: |
2022
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| Zugriff auf das übergeordnete Werk: | Journal of fish diseases |
| Schlagworte: | Journal Article RT-PCR RT-RPA VHSV rapid diagnostic RNA, Viral Recombinases |
| Zusammenfassung: | © 2022 John Wiley & Sons Ltd. Viral diseases of fish cause significant economic losses in the aquaculture industry. Viral haemorrhagic septicemia virus (VHSV) is one of the most important viral diseases that affects more than 80 fish species. Detection of the disease, especially in the field, is critical to managing disease prevention and control programmes. Recombinase polymerase amplification (RPA) is an isothermal method with a very short amplification period and a single incubation temperature ranging from 37 to 42°C, which is a good alternative to the polymerase chain reaction (PCR). This study aimed to develop an RPA assay as sensitive as a real-time RT-PCR to detect VHSV. For this purpose, primers and probes are designed for the same targeted region of gG of VHSV. The ssRNA standards were prepared to find the detection limits of the assay. Detection limits were found ten-fold differences between real-time RT-PCR and real-time RT-RPA. While the detection limit of the RT-PCR was found as 95.5 viral RNA molecules/reaction in 95% probit value, the detection limit of RT-RPA was found as 943.75 viral RNA molecules/reaction in 95% probit value using ssRNA standards. These results show that RPA is a suitable test for VHSV Ie detection |
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| Beschreibung: | Date Completed 14.07.2022 Date Revised 14.07.2022 published: Print-Electronic RefSeq: JF415086.1, JF415087.1, JF415088.1, JF415089.1, JF415090.1, JF415091.1, KM972678.1, KM972679.1, KM972680.1, LN877207.1, KM9729679.1 Citation Status MEDLINE |
| ISSN: | 1365-2761 |
| DOI: | 10.1111/jfd.13629 |