TY - JOUR
T1 - A novel array of real-time RT-PCR assays for the rapid pathotyping of type I avian paramyxovirus (APMV-1)
AU - Fortin, Andrea
AU - Laconi, Andrea
AU - Monne, Isabella
AU - Zohari, Siamak
AU - Andersson, Kristofer
AU - Grund, Christian
AU - Cecchinato, Mattia
AU - Crimaudo, Marika
AU - Valastro, Viviana
AU - D'Amico, Valeria
AU - Bortolami, Alessio
AU - Gastaldelli, Michele
AU - Varotto, Maria
AU - Abdelrahman, Amgad
AU - Amarin, Nadim
AU - Abubakar, Mustapha Bala
AU - Belayneh, Redeat
AU - Cyprien, Yapi Bokpè
AU - Christodoulou, Vasiliki
AU - Chvala, Ilya
AU - Dodovski, Aleksandar
AU - Ghafouri, Seyed Ali
AU - Giasuddin, Mohammed
AU - Hassan, Magdy
AU - Kammon, Abdulwahab
AU - Shittu, Ismaila
AU - Snoeck, Chantal J.
AU - Steensel, Mieke
AU - Suarez, David
AU - Torchetti, Mia Kim
AU - Tshipambe, Serge Mpiana
AU - Ouermi Zerbo, Lamouni Habibata
AU - Terregino, Calogero
AU - Panzarin, Valentina
AU - Newcastle Disease Collaborating Diagnostic Group
N1 - Funding Information:
This work was supported by the European Union within the framework of the activities foreseen by the EURL for AI/ND. The research was also partially funded by a grant from the European Commission (Development Cooperation Instruments) awarded to the project “EU Support to Livestock Disease Surveillance Knowledge Integration - LIDISKI ” ( FOOD/2019/410–957 ) within the frame of the Development of Smart Innovation through Research in Agriculture (DeSIRa) program and by the Italian Ministry of Health (Grant number RC IZSVe 05/2020 ).
Publisher Copyright:
© 2023 The Authors
PY - 2023/12
Y1 - 2023/12
N2 - Newcastle disease (ND) caused by virulent avian paramyxovirus type I (APMV-1) is a WOAH and EU listed disease affecting poultry worldwide. ND exhibits different clinical manifestations that may either be neurological, respiratory and/or gastrointestinal, accompanied by high mortality. In contrast, mild or subclinical forms are generally caused by lentogenic APMV-1 and are not subject to notification. The rapid discrimination of virulent and avirulent viruses is paramount to limit the spread of virulent APMV-1. The appropriateness of molecular methods for APMV-1 pathotyping is often hampered by the high genetic variability of these viruses that affects sensitivity and inclusivity. This work presents a new array of real-time RT-PCR (RT-qPCR) assays that enable the identification of virulent and avirulent viruses in dual mode, i.e., through pathotype-specific probes and subsequent Sanger sequencing of the amplification product. Validation was performed according to the WOAH recommendations. Performance indicators on sensitivity, specificity, repeatability and reproducibility yielded favourable results. Reproducibility highlighted the need for assays optimization whenever major changes are made to the procedure. Overall, the new RT-qPCRs showed its ability to detect and pathotype all tested APMV-1 genotypes and its suitability for routine use in clinical samples.
AB - Newcastle disease (ND) caused by virulent avian paramyxovirus type I (APMV-1) is a WOAH and EU listed disease affecting poultry worldwide. ND exhibits different clinical manifestations that may either be neurological, respiratory and/or gastrointestinal, accompanied by high mortality. In contrast, mild or subclinical forms are generally caused by lentogenic APMV-1 and are not subject to notification. The rapid discrimination of virulent and avirulent viruses is paramount to limit the spread of virulent APMV-1. The appropriateness of molecular methods for APMV-1 pathotyping is often hampered by the high genetic variability of these viruses that affects sensitivity and inclusivity. This work presents a new array of real-time RT-PCR (RT-qPCR) assays that enable the identification of virulent and avirulent viruses in dual mode, i.e., through pathotype-specific probes and subsequent Sanger sequencing of the amplification product. Validation was performed according to the WOAH recommendations. Performance indicators on sensitivity, specificity, repeatability and reproducibility yielded favourable results. Reproducibility highlighted the need for assays optimization whenever major changes are made to the procedure. Overall, the new RT-qPCRs showed its ability to detect and pathotype all tested APMV-1 genotypes and its suitability for routine use in clinical samples.
KW - Avian paramyxovirus type I (APMV-1)
KW - Avirulent
KW - Molecular pathotyping
KW - Newcastle disease
KW - Virulent
UR - http://www.scopus.com/inward/record.url?scp=85173994167&partnerID=8YFLogxK
UR - https://pubmed.ncbi.nlm.nih.gov/37722509
U2 - 10.1016/j.jviromet.2023.114813
DO - 10.1016/j.jviromet.2023.114813
M3 - Article
C2 - 37722509
SN - 0166-0934
VL - 322
JO - Journal of Virological Methods
JF - Journal of Virological Methods
M1 - 114813
ER -