Detection of equine herpesvirus type 1 using a real-time polymerase chain reaction

2006 ◽  
Vol 131 (1) ◽  
pp. 92-98 ◽  
Author(s):  
Ibrahim S. Diallo ◽  
Glen Hewitson ◽  
Lucia Wright ◽  
Barry J. Rodwell ◽  
Bruce G. Corney
1992 ◽  
Vol 38 (11) ◽  
pp. 1193-1196 ◽  
Author(s):  
Wania N. Wagner ◽  
Jaret Bogdan ◽  
Deborah Haines ◽  
Hugh G. G. Townsend ◽  
Vikram Misra

Although both equine herpesvirus type 1 (EHV-1) and equine herpesvirus type 4 (EHV-4) can be associated with respiratory disease, epizootics caused by EHV-1 are much more serious because the virus can cause abortions and paralysis. It is, therefore, important to identify the type of EHV involved in an outbreak by a test that is quick, sensitive, and reliable. We have adapted the polymerase chain reaction (PCR) to detect and distinguish between EHV-1 and EHV-4 in the same reaction. Primers for PCR were designed from the sequences of the glycoprotein B genes of EHV-1 and EHV-4. The PCR products derived from EHV-1 and EHV-4 were 135 and 326 base pairs, respectively, and could be readily separated by electrophoresis. The identity of the PCR products was confirmed by determining their nucleotide sequence, which agreed with the published sequence of the gB genes. The test could be performed directly on virus pelleted from small volumes (300 μL) of medium in which nasal swabs were transported and did not rely on the presence of infectious virus. The PCR was unaffected by conditions that reduced the infectivity of a virus preparation by 99%. The PCR detected EHV-4 in 5 of 10 nasal mucous samples taken from an outbreak of respiratory disease in race horses. Virus isolation in indicator cells was successful in detecting virus in four of the five samples positive by PCR. Key words: equine herpesvirus types 1 and 4, polymerase chain reaction, equine respiratory disease.


2000 ◽  
Vol 145 (9) ◽  
pp. 1773-1787 ◽  
Author(s):  
R. Carvalho ◽  
A. M. Oliveira ◽  
A. M. Souza ◽  
L. M. F. Passos ◽  
A. S. Martins

2006 ◽  
Vol 133 (1) ◽  
pp. 70-75 ◽  
Author(s):  
Gabriella Elia ◽  
Nicola Decaro ◽  
Vito Martella ◽  
Marco Campolo ◽  
Costantina Desario ◽  
...  

2015 ◽  
Vol 18 (4) ◽  
pp. 833-839 ◽  
Author(s):  
K. Stasiak ◽  
J. Rola ◽  
J.F. Zmudzinski

Abstract A highly sensitive and specific real-time PCR assay was used for detection and quantitation of equine herpesvirus type 1 (EHV-1) in the different internal organs of aborted fetuses. Tissue samples from 23 aborted fetuses submitted to the Department of Virology of the National Veterinary Research Institute in Pulawy between 2012 and 2013 were used for testing. Total DNA was extracted using a phenol-chloroform-isoamyl alcohol standard protocol. A real-time PCR with forward and reverse primers encompassing a highly conserved region encoding viral glycoprotein B was adapted for diagnosis of EHV-1 infection. The detection limit of the assay was shown to be 6.0×100 of viral DNA copies and the obtained standard curve exhibited a linear range from 100 to 107 molecules. Sixteen out of twenty three aborted fetuses (69.5%) were positive for EHV-1 in real-time PCR. The highest EHV-1 DNA load was obtained for liver (mean Ct value: 15.7) and lung (18.2) samples, while the lowest was in the thymus (29.6) and placenta (28.4).


2012 ◽  
Vol 15 (1) ◽  
pp. 151-153 ◽  
Author(s):  
A. Golke ◽  
J. Cymerys ◽  
A. Słońska ◽  
T. Dzieciątkowski ◽  
A. Chmielewska ◽  
...  

The xCELLigence system for real-time and label-free analysis of neuronal and dermal cell response to Equine Herpesvirus type 1 infection Real-time cell electronic sensing (RT-CES) based on impedance measurements is an emerging technology for analyzing the status of cells in vitro. It allows label-free, real time monitoring of the biological status of cells. The present study was designed to assess dynamic data on the cell processes during equine herpesvirus type 1 (EHV-1) infection of ED (equine dermal) cells and primary murine neuronal cell culture. We have demonstrated that the xCELLigence system with dynamic monitoring can be used as a rapid diagnostic tool both to analyze cellular behavior and to investigate the effect of viral infection.


Vaccine ◽  
2008 ◽  
Vol 26 (19) ◽  
pp. 2335-2343 ◽  
Author(s):  
Cristina Rosas ◽  
Gerlinde R. Van de Walle ◽  
Stephan M. Metzger ◽  
Karin Hoelzer ◽  
Edward J. Dubovi ◽  
...  

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