tetano
Editor, Senior Moderator
Viruses
. 2022 Feb 17;14(2):415.
doi: 10.3390/v14020415.
Improved Subtyping of Avian Influenza Viruses Using an RT-qPCR-Based Low Density Array: 'Riems Influenza a Typing Array', Version 2 (RITA-2)
Kareem E Hassan[SUP] 1 2 [/SUP], Ann Kathrin Ahrens[SUP] 1 [/SUP], Ahmed Ali[SUP] 2 [/SUP], Magdy F El-Kady[SUP] 2 [/SUP], Hafez M Hafez[SUP] 3 [/SUP], Thomas C Mettenleiter[SUP] 4 [/SUP], Martin Beer[SUP] 1 [/SUP], Timm Harder[SUP] 1 [/SUP]
Affiliations
Abstract
Avian influenza virus (AIV) variants emerge frequently, which challenges rapid diagnosis. Appropriate diagnosis reaching the sub- and pathotype level is the basis of combatting notifiable AIV infections. Real-time RT-PCR (RT-qPCR) has become a standard diagnostic tool. Here, a total of 24 arrayed RT-qPCRs is introduced for full subtyping of 16 hemagglutinin and nine neuraminidase subtypes of AIV. This array, designated Riems Influenza A Typing Array version 2 (RITA-2), represents an updated and economized version of the RITA-1 array previously published by Hoffmann et al. RITA-2 provides improved integration of assays (24 instead of 32 parallel reactions) and reduced assay volume (12.5 µL). The technique also adds RT-qPCRs to detect Newcastle Disease (NDV) and Infectious Bronchitis viruses (IBV). In addition, it maximizes inclusivity (all sequences within one subtype) and exclusivity (no intersubtypic cross-reactions) as shown in validation runs using a panel of 428 AIV reference isolates, 15 reference samples each of NDV and IBV, and 122 clinical samples. The open format of RITA-2 is particularly tailored to subtyping influenza A virus of avian hosts and Eurasian geographic origin. Decoupling and re-arranging selected RT-qPCRs to detect specific AIV variants causing epizootic outbreaks with a temporal and/or geographic restriction is possible.
Keywords: Newcastle disease virus; avian influenza; diagnosis; infectious bronchitis virus; real-time RT-PCR.
. 2022 Feb 17;14(2):415.
doi: 10.3390/v14020415.
Improved Subtyping of Avian Influenza Viruses Using an RT-qPCR-Based Low Density Array: 'Riems Influenza a Typing Array', Version 2 (RITA-2)
Kareem E Hassan[SUP] 1 2 [/SUP], Ann Kathrin Ahrens[SUP] 1 [/SUP], Ahmed Ali[SUP] 2 [/SUP], Magdy F El-Kady[SUP] 2 [/SUP], Hafez M Hafez[SUP] 3 [/SUP], Thomas C Mettenleiter[SUP] 4 [/SUP], Martin Beer[SUP] 1 [/SUP], Timm Harder[SUP] 1 [/SUP]
Affiliations
- PMID: 35216008
- PMCID: PMC8879595
- DOI: 10.3390/v14020415
Abstract
Avian influenza virus (AIV) variants emerge frequently, which challenges rapid diagnosis. Appropriate diagnosis reaching the sub- and pathotype level is the basis of combatting notifiable AIV infections. Real-time RT-PCR (RT-qPCR) has become a standard diagnostic tool. Here, a total of 24 arrayed RT-qPCRs is introduced for full subtyping of 16 hemagglutinin and nine neuraminidase subtypes of AIV. This array, designated Riems Influenza A Typing Array version 2 (RITA-2), represents an updated and economized version of the RITA-1 array previously published by Hoffmann et al. RITA-2 provides improved integration of assays (24 instead of 32 parallel reactions) and reduced assay volume (12.5 µL). The technique also adds RT-qPCRs to detect Newcastle Disease (NDV) and Infectious Bronchitis viruses (IBV). In addition, it maximizes inclusivity (all sequences within one subtype) and exclusivity (no intersubtypic cross-reactions) as shown in validation runs using a panel of 428 AIV reference isolates, 15 reference samples each of NDV and IBV, and 122 clinical samples. The open format of RITA-2 is particularly tailored to subtyping influenza A virus of avian hosts and Eurasian geographic origin. Decoupling and re-arranging selected RT-qPCRs to detect specific AIV variants causing epizootic outbreaks with a temporal and/or geographic restriction is possible.
Keywords: Newcastle disease virus; avian influenza; diagnosis; infectious bronchitis virus; real-time RT-PCR.