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

Lu, Zhengchun; Dubovi, Edward J.; Zylich, Nancy C.; Crawford, P. Cynda; Sells, Stephen F.; Go, Yun Young; Loynachan, Alan T.; Timoney, Peter J.; Chambers, Thomas M.; Balasuriya, Udeni B. R.

doi:10.1177/104063871002200614

Cited by 13 publications

(9 citation statements)

References 20 publications

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“…To construct the complementary DNA, the reverse-transcriptase SuperScriptII (Invitrogen) enzyme was used. Real-time RT-PCR was used with primers addressed to a fragment of the gene NP, according to Lu et al 22 To identify the viral RNA of human influenza A, the primers and probe were identical to those used for the real-time RT-PCR previously described for gene M of most influenza A subtypes. 23 Primers used in this research were designed to align most type A influenza viruses originating from multiple species.…”

Section: Detection Of Viral Rnamentioning

confidence: 99%

Evidence of transmission and risk factors for influenza A virus in household dogs and their owners

Ramírez‐Martínez¹,

Contreras‐Luna²,

Luz³

et al. 2013

Influenza Resp Viruses

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BackgroundThe possible transmission of influenza A virus between dogs and humans is important, as in Mexico City there are approximately 1·2 million dogs. We present the first evidence of influenza A virus infection in household dogs in Mexico.ObjectivesThe objective of this study was to identify the presence of antibodies against influenza A virus in dogs and their owners, as well as the presence of RNA of influenza A virus in nasal exudates of dogs and, thereby, assess the possible transmission of the virus between humans and dogs.MethodsSerum samples from household dogs and their owners were analyzed to detect the presence of antibodies against three subtypes of human influenza virus (H1N1pdm09, H1N1, and H3N2), as well as subtype H3N8 of equine influenza. We analyzed dog nasal exudates to detect influenza viral RNA. The relationship between the seropositivity of dogs and various factors (age, sex, constantly at home, and seropositivity of owners) was statistically analyzed.ResultsSeroprevalence for human influenza in dogs was 0·9% (1 of 113), and it was 4% (5 of 113) for equine influenza. In humans, seroprevalence was 22% for subtype H1N1pdm09, 20% for subtype H1N1, and 11% for subtype H3N2. No significant association (P > 0·05) was found between seropositivity and any of the assessed factors. Furthermore, no viral RNA was detected in the nasal exudate samples.ConclusionsResults revealed seroprevalence of the influenza virus in household dogs in Mexico City. It can be assumed that dogs are currently becoming infected with different subtypes of influenza viruses.

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Section: Detection Of Viral Rnamentioning

confidence: 99%

Evidence of transmission and risk factors for influenza A virus in household dogs and their owners

Ramírez‐Martínez¹,

Contreras‐Luna²,

Luz³

et al. 2013

Influenza Resp Viruses

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“…Particularly, virological and serological surveys have documented that H1N1/2009, cH3N2, hH3N2, and equine-origin H3N8 influenza viruses are consistently circulating in dogs [3–6]. A few methods for detecting equine-origin H3N8 and cH3N2 CIV subtypes have been developed [30–32]. However, no study has described a method for distinguishing different influenza virus subtypes that circulate in dogs.…”

Section: Discussionmentioning

confidence: 99%

A Multiplex RT-PCR Assay for Detection and Differentiation of Avian-Origin Canine H3N2, Equine-Origin H3N8, Human-Origin H3N2, and H1N1/2009 Canine Influenza Viruses

Wang

et al. 2017

PLoS ONE

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Virological and serological surveys have documented that H1N1/2009, avian-origin canine H3N2 (cH3N2), seasonal human-origin H3N2 (hH3N2), and equine-origin H3N8 influenza viruses are consistently circulating in dogs. In the present study, a multiplex reverse-transcriptase polymerase chain reaction (mRT-PCR) assay was developed for simultaneous detection and differentiation of these influenza viruses. Four primer sets were designed to target the hemagglutinin genes of H1N1/2009, cH3N2, hH3N2, and H3N8 canine influenza viruses (CIVs). This mRT-PCR assay demonstrated high specificity and sensitivity for the four CIV subtypes. Additionally, mRT-PCR results obtained from 420 clinical samples were consistent with those obtained by the conventional virus isolation method. Our mRT-PCR assay is reliable for clinical diagnosis and rapid identification of CIVs.

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“…These primers were tested to verify reactivity with the specific AIV studied in ponies. RNA copy numbers were calculated against a standard curve generated using in vitro ‐transcribed RNA made from cloned M1 cDNA as described . Sera were obtained at Day 0 and again at Day 14 post‐infection, and virus‐specific serum antibodies were measured using the hemagglutination‐inhibition (HI) assay using both ether‐treated and untreated virus antigens.…”

Section: Avian Influenza Viruses Testedmentioning

confidence: 99%

“…RNA copy numbers were calculated against a standard curve generated using in vitro-transcribed RNA made from cloned M1 cDNA as described. 14 Results of these pony infection experiments were almost all negative. No pony exhibited disease signs or seroconverted to the AIV used for infection.…”

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confidence: 96%

Replication of avian influenza viruses in equine tracheal epithelium but not in horses

Chambers

Balasuriya

Reedy

et al. 2013

Influenza Resp Viruses

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We evaluated a hypothesis that horses are susceptible to avian influenza viruses by in vitro testing, using explanted equine tracheal epithelial cultures, and in vivo testing by aerosol inoculation of ponies. Results showed that several subtypes of avian influenza viruses detectably replicated in vitro. Three viruses with high in vitro replication competence were administered to ponies. None of the three demonstrably replicated or caused disease signs in ponies. While these results do not exhaustively test our hypothesis, they do highlight that the tracheal explant culture system is a poor predictor of in vivo infectivity.

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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

Abstract: Abstract. 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

Cited by 13 publications

References 20 publications

Evidence of transmission and risk factors for influenza A virus in household dogs and their owners

Evidence of transmission and risk factors for influenza A virus in household dogs and their owners

A Multiplex RT-PCR Assay for Detection and Differentiation of Avian-Origin Canine H3N2, Equine-Origin H3N8, Human-Origin H3N2, and H1N1/2009 Canine Influenza Viruses

Replication of avian influenza viruses in equine tracheal epithelium but not in horses

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