A real-time PCR assay for measuring alcelaphine herpesvirus-1 DNA

Alcelaphine herpesvirus-1 (AlHV-1) is a rhadinovirus that causes malignant catarrhal fever in certain ruminant species and is an important pathogen in Africa and other areas where carrier species and clinically susceptible ruminants intermingle. In this study, a real-time PCR for AlHV-1 DNA was deve...

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Bibliographic Details
Published in:Journal of virological methods 2005-11, Vol.129 (2), p.186-190
Main Authors: Traul, Donald L., Elias, Shirley, Taus, Naomi S., Herrmann, Lynn M., Oaks, J. Lindsay, Li, Hong
Format: Article
Language:English
Subjects:
Animals
Biological and medical sciences
Cattle
Cattle Diseases - diagnosis
Cattle Diseases - virology
Fundamental and applied biological sciences. Psychology
Gammaherpesvirinae - genetics
Gammaherpesvirinae - isolation & purification
Herpesviridae Infections - diagnosis
Herpesviridae Infections - veterinary
Microbiology
Polymerase Chain Reaction - methods
Reproducibility of Results
Rhadinovirus
Ruminantia
Sensitivity and Specificity
Techniques used in virology
Tumor Virus Infections - diagnosis
Tumor Virus Infections - veterinary
Tumor Virus Infections - virology
Viral Structural Proteins - genetics
Virology
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Summary:Alcelaphine herpesvirus-1 (AlHV-1) is a rhadinovirus that causes malignant catarrhal fever in certain ruminant species and is an important pathogen in Africa and other areas where carrier species and clinically susceptible ruminants intermingle. In this study, a real-time PCR for AlHV-1 DNA was developed and compared to an established nested PCR. The nested PCR amplifies a region of the AlHV-1 gene coding for a transactivator protein (ORF 50), while the real-time PCR assay targets the AlHV-1 gene coding for a tegument protein (ORF 3). The real-time PCR assay reproducibly detected 10 copies of target DNA. In a dilution series of the target DNA there was linearity of the assay across 8 orders of magnitude (10 1–10 9 copies). The nested PCR was more sensitive (≅1 log) than the real-time PCR. The assay specifically amplified samples containing only AlHV-1, but not other common herpesviruses of cattle. In conclusion, we have developed a rapid, relatively sensitive, and reliable real-time PCR assay specific for AlHV-1. Similar to the real-time PCR for Ovine herpesvirus-2, this assay should prove useful for differential diagnostics of clinical MCF and for research to better define the epidemiology of AlHV-1 in wildebeest as well as in animals with wildebeest-associated MCF.
ISSN:0166-0934
1879-0984
DOI:10.1016/j.jviromet.2005.05.021