TY - JOUR
T1 - Diagnostic application of H3N8-specific equine influenza real-time reverse transcription polymerase chain reaction assays for the detection of canine influenza virus in clinical specimens
AU - Lu, Zhengchun
AU - Dubovi, Edward J.
AU - Zylich, Nancy C.
AU - Crawford, P. Cynda
AU - Sells, Stephen
AU - Go, Yun Young
AU - Loynachan, Alan T.
AU - Timoney, Peter J.
AU - Chambers, Thomas M.
AU - Balasuriya, Udeni B.R.
N1 - Copyright:
Copyright 2017 Elsevier B.V., All rights reserved.
PY - 2010/11
Y1 - 2010/11
N2 - The objective of the current study was to determine the capability of 3 recently described onestep TaqMan real time reverse transcription polymerase chain reaction (real-time RT-PCR) assays targeting the nucleoprotein (NP), matrix (M), and hemagglutinin (HA) genes of H3N8 Equine influenza virus (EIV NP, EIV M, and EIV HA3 assays, respectively) to detect Canine influenza virus (CIV). The assays were initially evaluated with nucleic acid extracted from tissue culture fluid (TCF) containing the A/canine/FL/43/04 strain of Influenza A virus associated with the 2004 canine influenza outbreak in Florida. The EIV NP, EIV M, and EIV HA3 assays could detect CIV nucleic acid at threshold cycle (Ct) values of 16.31, 23.71, and 15.28, respectively. Three assays using TCF or allantoic fluid (AF) samples containing CIV (n 5 13) and archived canine nasal swab samples (n 5 20) originally submitted for laboratory diagnosis of CIV were further evaluated. All TCF and AF samples, together with 10 nasal swab samples that previously tested positive fo virus by attempted isolation in embryonated hens' eggs or Madin-Darby canine kidney cells, were positive in all 3 real-time RT-PCR assays. None of the 3 assays detected the H1N1 Swine influenza virus strain in current circulation. These findings demonstrate that previously described real-time RT-PCR assays targeting NP, M, and H3 HA gene segments of H3N8 EIV are also valuable for the diagnosis of CIV infection in dogs. The assays could expedite the detection and identification of CIV.
AB - The objective of the current study was to determine the capability of 3 recently described onestep TaqMan real time reverse transcription polymerase chain reaction (real-time RT-PCR) assays targeting the nucleoprotein (NP), matrix (M), and hemagglutinin (HA) genes of H3N8 Equine influenza virus (EIV NP, EIV M, and EIV HA3 assays, respectively) to detect Canine influenza virus (CIV). The assays were initially evaluated with nucleic acid extracted from tissue culture fluid (TCF) containing the A/canine/FL/43/04 strain of Influenza A virus associated with the 2004 canine influenza outbreak in Florida. The EIV NP, EIV M, and EIV HA3 assays could detect CIV nucleic acid at threshold cycle (Ct) values of 16.31, 23.71, and 15.28, respectively. Three assays using TCF or allantoic fluid (AF) samples containing CIV (n 5 13) and archived canine nasal swab samples (n 5 20) originally submitted for laboratory diagnosis of CIV were further evaluated. All TCF and AF samples, together with 10 nasal swab samples that previously tested positive fo virus by attempted isolation in embryonated hens' eggs or Madin-Darby canine kidney cells, were positive in all 3 real-time RT-PCR assays. None of the 3 assays detected the H1N1 Swine influenza virus strain in current circulation. These findings demonstrate that previously described real-time RT-PCR assays targeting NP, M, and H3 HA gene segments of H3N8 EIV are also valuable for the diagnosis of CIV infection in dogs. The assays could expedite the detection and identification of CIV.
KW - Canine influenza
KW - Diagnosis
KW - Dogs
KW - Equine influenza virus-2 (H3N8)
KW - Real-time reverse transcription polymerase chain reaction
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U2 - 10.1177/104063871002200614
DO - 10.1177/104063871002200614
M3 - Article
C2 - 21088179
AN - SCOPUS:78649582476
SN - 1040-6387
VL - 22
SP - 942
EP - 945
JO - Journal of Veterinary Diagnostic Investigation
JF - Journal of Veterinary Diagnostic Investigation
IS - 6
ER -