Arrêt de service programmé du vendredi 10 juin 16h jusqu’au lundi 13 juin 9h. Pour en savoir plus
Accéder directement au contenu Accéder directement à la navigation
Article dans une revue

One-step cross-genogroup multiplex RT-qPCR with an internal control system for the detection of infectious pancreatic necrosis virus (IPNV)

Abstract : Infectious pancreatic necrosis virus (IPNV) causes great losses in fish hatcheries world-wide. The detection of IPNV can be challenging in certain circumstances, particularly due to low viral load and the genetic variability of this RNA virus. For the first time, this project created a quantitative triplex real-time reverse transcription PCR (RT-qPCR), including an endogenous control system, for specific, sensitive and rapid detection of IPNV in routine diagnostics. Multiple sequence alignment of 46 nucleotide sequences of the segment A genome obtained from the NCBI database allowed the design of two RT-qPCR systems covering the IPNV genogroup 1 and genogroups 2-5, respectively. The completed triplex RT-qPCR including a salmonid-specific endogenous control showed high Specificity and an analytical sensitivity of 20-40 oligonucleotide copies. Testing of dilution series of virus-loaded cell culture suspensions proved equality of the triplex RT-qPCR with virus detection in cell culture and a higher sensitivity than conventional RT-PCR in field samples. In comparative studies of a total of 77 field samples tested, 51 showed identical positive and 19 identical negative results in cell culture and the triplex RT-qPCR. However, seven other samples yielded positive results in the triplex RT-qPCR, but negative results in cell culture.
Type de document :
Article dans une revue
Liste complète des métadonnées

https://hal-u-picardie.archives-ouvertes.fr/hal-03548807
Contributeur : Louise Dessaivre Connectez-vous pour contacter le contributeur
Soumis le : lundi 31 janvier 2022 - 09:30:21
Dernière modification le : vendredi 25 mars 2022 - 18:12:15

Identifiants

Collections

Citation

Marc Hoferer, Anne Braun, Julia Skrypski, Sabine Bock, Sabine Thalheim, et al.. One-step cross-genogroup multiplex RT-qPCR with an internal control system for the detection of infectious pancreatic necrosis virus (IPNV). JOURNAL OF VIROLOGICAL METHODS, 2017, 247, pp.68-76. ⟨10.1016/j.jviromet.2017.05.015⟩. ⟨hal-03548807⟩

Partager

Métriques

Consultations de la notice

3