Detection of Equine Herpesvirus 3 in Equine Skin Lesions by Polymerase Chain Reaction

Paraffin-embedded organ samples from 28 aborted fetuses and three foals, partly archival and partly sampled in 1991, were examined by polymerase chain reaction (PCR) and immunohistochemistry for the presence of DNA and antigens, respectively, specific for equine herpesvirus 1 (EHV- 1). Virologic examination had been performed on 23 of the aborted fetuses. DNA fragments specific for EHV-1 were identified by PCR, and EHV-1 antigens were identified in situ by immunohistochemistry, with an agreement between the methods of 94% (kappa = 0.85). Compared with virus isolation, PCR agreement was 87% (kappa 0.69), and IH agreement was 82% (kappa = 0.47). These results showed that there was moderate to almost perfect agreement among the different methods and that PCR and immunohistochemistry are powerful tools for the identification of EHV-1 in paraffin-embedded tissues. The techniques give more rapid results than virus isolation and also detect inactivated virus, which are not identified by standard virus isolation. These techniques also make retrospective studies possible.

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Detection of equine herpesvirus and differentiation of equine herpesvirus type 1 from type 4 by the polymerase chain reaction

Canadian Journal of Microbiology ◽

10.1139/m92-196 ◽

1992 ◽

Vol 38 (11) ◽

pp. 1193-1196 ◽

Cited By ~ 31

Author(s):

Wania N. Wagner ◽

Jaret Bogdan ◽

Deborah Haines ◽

Hugh G. G. Townsend ◽

Vikram Misra

Keyword(s):

Polymerase Chain Reaction ◽

Respiratory Disease ◽

Equine Herpesvirus ◽

Chain Reaction ◽

Herpesvirus Type ◽

Equine Herpesvirus Type ◽

Pcr Products ◽

Equine Herpesvirus Type 1 ◽

Polymerase Chain

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.

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Molecular Detection of Equine Herpesvirus Types 1 and 4 Infection in Healthy Horses in Isfahan Central and Shahrekord Southwest Regions, Iran

BioMed Research International ◽

10.1155/2015/917854 ◽

2015 ◽

Vol 2015 ◽

pp. 1-7 ◽

Cited By ~ 3

Author(s):

Taghi Taktaz Hafshejani ◽

Shahin Nekoei ◽

Behnam Vazirian ◽

Abbas Doosti ◽

Faham Khamesipour ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

Molecular Detection ◽

Equine Herpesvirus ◽

Chain Reaction ◽

Blood Samples ◽

Thoroughbred Horses ◽

Polymerase Chain

This study was undertaken to investigate molecularly the occurrence of EHV-1 and EHV-4 infection among equine population in regions, Iran. Blood samples from 53 and 37 randomly selected horses settled in Isfahan and Shahrekord, Iran, respectively, were collected. Detection of EHV-1 and EHV-4 genes in the blood samples was done using polymerase chain reaction (PCR). Out of 53 and 37 samples from Isfahan and Shahrekord, 4 (18.18%) and 3 (8.10%) were positive for PCR of EHV-1, respectively. Nine (16.98%) and 6 (16.21%) were positive for PCR of EHV-4, while 6 (11.32%) and 3 (8.10%) were positive for PCR of both EHV-1 and EHV-4, in Isfahan and Shahrekord, respectively. Of the 7 blood samples positive for EHV-1, 4 (16.66%) and 3 (8.10%) were from horses >3 years old while 2 (18.18%) and 1 (16.66%) were from 2-3 years old horses, in Isfahan and Shahrekord, respectively. Out of the 7 and 3 samples positive for PCR of EHV-1 in Isfahan and Shahrekord, 4 (22.2%) and 1 (7.69%) were Standardbred, while 3 (14.28%) and 2 (13.33%) were Thoroughbreds, respectively. EHV-4 was detected in blood of 4 (22.22%) and 2 (15.83%) Standardbreds and from 4 (19.04%) and 4 (26.66%) Thoroughbred horses in Isfahan and Shahrekord, respectively. This study has shown that horses settled in Isfahan central and Shahrekord southwest regions, Iran, are infected by EHV-1 and EHV-4 and thus serve as potential reservoirs and disseminators of the viruses.

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