Detection of Equine Herpesvirus-1 in Nasal Swabs of Horses by Quantitative Real-Time PCR

Upper respiratory disease has been a serious problem in Standardbred horses on racetracks in Ontario, with outbreaks occurring once or twice annually in late winter and early spring seasons. To determine the causes of these epidemics, a 3-year investigation was carried out in which nasal swabs and serum samples were obtained at intervals from apparently healthy horses and from horses suffering from upper respiratory disease. The nasal swabs were used to isolate bacteria and viruses. The serum samples were examined for the presence and level of antibodies to equine influenza viruses and equine herpesvirus 1. None of the bacteria isolated were associated with the outbreaks of disease. Equine herpesvirus 2 was isolated 72 times from both diseased and apparently healthy horses. Equine herpesvirus 1 was isolated 10 times from horses with respiratory disease, both during and between epidemics. Influenza equine/1 virus was isolated seven times and influenza equine/2 was isolated once during severe outbreaks of upper respiratory disease. Serological evidence confirmed that influenza viruses were the causes of the major epidemics, with the equine/1 strain being involved most often.

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Development of a Real-Time Polymerase Chain Reaction Assay for Rapid Diagnosis of Neuropathogenic Strains of Equine Herpesvirus-1

Journal of Veterinary Diagnostic Investigation ◽

10.1177/104063870701900110 ◽

2007 ◽

Vol 19 (1) ◽

pp. 69-72 ◽

Cited By ~ 36

Author(s):

George P. Allen

Keyword(s):

Polymerase Chain Reaction ◽

Real Time ◽

Polymerase Chain Reaction Assay ◽

Rapid Diagnosis ◽

Equine Herpesvirus ◽

Chain Reaction ◽

Equine Herpesvirus 1 ◽

Herpesvirus 1 ◽

Polymerase Chain

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Establishment and Application of a Taqman Reverse Transcriptase Quantitative Real Time Pcr Assay for Feline Calicivirus

10.21203/rs.3.rs-267291/v1 ◽

2021 ◽

Author(s):

jingjie zhao ◽

Lin Liang ◽

Guangzhi Zhang ◽

Wenhui Li ◽

Shaohan Li ◽

...

Keyword(s):

Reverse Transcriptase ◽

Real Time ◽

Real Time Pcr ◽

Limit Of Detection ◽

Clinical Samples ◽

Feline Calicivirus ◽

Quantitative Real Time Pcr ◽

Pcr Assay ◽

Virus Content ◽

Nasal Swabs

Abstract Feline calicivirus (FCV) is an infectious pathogen that causes disease in cats. With the current emergence of FCV-associated virulent systemic disease (FCV VSD) worldwide, the establishment of a rapid, sensitive, and reproducible diagnostic assay for its detection is important to inform prevention and control strategies. In this study, specific primers and TaqMan-FAM probes were designed based on the conserved regions of the FCV genome sequence, and a TaqMan reverse transcriptase quantitative real time PCR assay was established. This assay could specifically detected the FCV genome. The assay had a wide dynamic range, with linear detection in the range of 9.6×109 copies/μL to 9.6×100 copies/μL, with a limit of detection of 9.6×100 copies/μL, showing high sensitivity and repeatability. In addition, we used this assay to evaluated clinical samples (n=100) taken from cats from across China for the presence/absence of FCV genetic material For samples with low virus content, the positive detection rate of TaqMan reverse transcriptase quantitative real time PCR assay (RT-qPCR) was much higher than that of conventional reverse transcriptase PCR assay (cRT-PCR). And The qRT-PCR assay was used to detect the viral load of cat swabs within 17 days after FCV infection. From days 1-9, the oral and nasal swabs generally had higher viral loads than the anal swabs. While from days 10-17, the levels in the oral and nasal swabs being generally lower than those in the anal swabs. Overall, this FCV TaqMan RT-qPCR assay assay represents a rapid and accurate.

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