tetano
Editor, Senior Moderator
J Virol Methods
. 2022 Aug 3;308:114594.
doi: 10.1016/j.jviromet.2022.114594. Online ahead of print.
Evaluation of PCR-based hemagglutinin subtyping as a tool to aid in surveillance of avian influenza viruses in migratory wild birds
Shahan Azeem[SUP] 1 [/SUP], Baoqing Guo[SUP] 2 [/SUP], Dong Sun[SUP] 3 [/SUP], Mary L Killian[SUP] 4 [/SUP], John A Baroch[SUP] 5 [/SUP], Kyoung-Jin Yoon[SUP] 6 [/SUP]
Affiliations
Abstract
The surveillance of migratory wild birds (MWBs) for avian influenza virus (AIV) allows detecting the emergence of highly pathogenic AIV that can infect domestic poultry and mammals, new subtypes, and antigenic/genetic variants. The current AIV surveillance system for MWBs in the United States is based on virus isolation (VI) followed by sequencing isolates. This system primarily focuses on the early detection of H5 and H7 AIVs. However, it is suboptimal in assessing diverse AIV subtypes at any given time because of the low VI success rate. To improve such a shortfall, a SYBR® Green-based real-time reverse transcription-polymerase chain reaction (rtRT-PCR) panel was developed for direct HA subtyping of AIVs in oropharyngeal-cloacal (OPC) swabs from MWBs. Under optimal conditions, the PCR panel detected AIVs of all 16 different HA subtypes with an average limit of detection of 10[SUP]2.6[/SUP] copies/reaction (2 μl of extract). In testing 90 OPC swabs from 13 MWB species, the PCR provided a significantly faster turnaround of results and demonstrated the presence of more subtypes and concurrent infection among MWBs compared to what the current surveillance testing algorithm showed. In conclusion, newly developed SYBR® Green rtRT-PCR panel can be a useful tool for monitoring MWBs for AIVs.
Keywords: Avian influenza virus; Migratory wild birds; Oropharyngeal-cloacal swab; Polymerase chain reaction; SYBR® Green; Subtyping; Virus isolation.
. 2022 Aug 3;308:114594.
doi: 10.1016/j.jviromet.2022.114594. Online ahead of print.
Evaluation of PCR-based hemagglutinin subtyping as a tool to aid in surveillance of avian influenza viruses in migratory wild birds
Shahan Azeem[SUP] 1 [/SUP], Baoqing Guo[SUP] 2 [/SUP], Dong Sun[SUP] 3 [/SUP], Mary L Killian[SUP] 4 [/SUP], John A Baroch[SUP] 5 [/SUP], Kyoung-Jin Yoon[SUP] 6 [/SUP]
Affiliations
- PMID: 35931229
- DOI: 10.1016/j.jviromet.2022.114594
Abstract
The surveillance of migratory wild birds (MWBs) for avian influenza virus (AIV) allows detecting the emergence of highly pathogenic AIV that can infect domestic poultry and mammals, new subtypes, and antigenic/genetic variants. The current AIV surveillance system for MWBs in the United States is based on virus isolation (VI) followed by sequencing isolates. This system primarily focuses on the early detection of H5 and H7 AIVs. However, it is suboptimal in assessing diverse AIV subtypes at any given time because of the low VI success rate. To improve such a shortfall, a SYBR® Green-based real-time reverse transcription-polymerase chain reaction (rtRT-PCR) panel was developed for direct HA subtyping of AIVs in oropharyngeal-cloacal (OPC) swabs from MWBs. Under optimal conditions, the PCR panel detected AIVs of all 16 different HA subtypes with an average limit of detection of 10[SUP]2.6[/SUP] copies/reaction (2 μl of extract). In testing 90 OPC swabs from 13 MWB species, the PCR provided a significantly faster turnaround of results and demonstrated the presence of more subtypes and concurrent infection among MWBs compared to what the current surveillance testing algorithm showed. In conclusion, newly developed SYBR® Green rtRT-PCR panel can be a useful tool for monitoring MWBs for AIVs.
Keywords: Avian influenza virus; Migratory wild birds; Oropharyngeal-cloacal swab; Polymerase chain reaction; SYBR® Green; Subtyping; Virus isolation.