Journal article
Analytical validation of one-step realtime RT-PCR for detection of infectious hematopoietic necrosis virus (IHNV)
Infectious hematopoietic necrosis virus (IHNV) is the causative agent of infectious hematopoietic necrosis, a disease listed by both the European Union (EU) and the World Organisation for Animal Health (OIE). Recently, molecular methods based on real-time RT-PCR have been approved for surveillance of IHNV within the EU, using the two-step real-time RT-PCR developed by Purcell et al. (2013).
The two-step procedure has some disadvantages compared to one-step methods due to the higher risk of contamination and it being a more laborious test. Here we show that modifying the original protocol to one-step real-time RT-PCR does not significantly compromise the analytical parameters of the method and it provides a similar detection limit for detection of IHNV when compared with cell culture methods.
The modified one-step protocol is robust and reproducible as shown by an inter-laboratory proficiency test carried out in eight laboratories in Europe.
Language: | English |
---|---|
Year: | 2020 |
Pages: | 261-272 |
ISSN: | 10249494 and 01080288 |
Types: | Journal article |
ORCIDs: | Cuenca, A. , Vendramin, N. and Olesen, N. Jørgen |