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United States Department of Agriculture

Agricultural Research Service

Title: A Real-Time Pcr Assay for Measuring Alcelaphine Herpesvirus-1 DNA

Authors
item Traul, Donald
item Elias, Shirley - WASHINGTON STATE UNIV.
item Taus, Naomi
item Hoesing, Lynn
item Oaks, Lindsay - WASHINGTON STATE UNIV.
item Li, Hong

Submitted to: Journal of Virological Methods
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: May 31, 2005
Publication Date: July 5, 2005
Citation: Traul, D., Elias, S., Taus, N.S., Hoesing, L.M., Oaks, L.J., Li, H. 2005. A real-time PCR assay for measuring alcelaphine herpesvirus-1 DNA. Journal of Virological Methods. 129:186-190.

Interpretive Summary: Alcelaphine herpesvirus 1 (AlHV-1) is a virus that causes malignant catarrhal fever and is an important pathogen in Africa and other areas where carrier species and clinically susceptible ruminants intermingle. We developed an improved quantitative diagnostic assay for detecting viral DNA using real-time polymerase chain reaction (PCR). The real-time PCR assay could detect as little as 10 copies of viral DNA with linearity across 8 orders of magnitude. The assay specifically detected samples containing only AlHV-1, but not other members of the MCF virus group including: Ovine herpesvirus 2, Alcelaphine herpesvirus 2, Caprine herpesvirus 2-the virus causing clinical MCF of unknown origin, or three recently reported MCF viruses found in muskox, ibex and oryx. The assay did not detect other common herpesviruses of cattle, including: Bovine herpesvirus 1, Bovine herpesvirus 4, or Bovine lymphotropic herpesvirus 1. This assay should be valuable for improved diagnostic and quantitative data of animals infected with AlHV-1.

Technical Abstract: Alcelaphine herpesvirus 1 (AlHV-1) causes malignant catarrhal fever and is an important pathogen in Africa and other areas where carrier species and clinically susceptible ruminants intermingle. In this study, a real-time quantitative PCR for AlHV-1 DNA was developed and compared to an established nested PCR. The nested PCR amplifies 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 with linearity across 8 orders of magnitude. 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 members of the MCF virus group including: Ovine herpesvirus 2, Alcelaphine herpesvirus 2, Caprine herpesvirus 2-the virus causing clinical MCF of unknown origin, or three recently reported MCF viruses found in muskox, ibex and oryx. The assay did not amplify other common herpesviruses of cattle, including: Bovine herpesvirus 1, Bovine herpesvirus 4, or Bovine lymphotropic herpesvirus 1. This assay should be valuable for rapid diagnostic and quantitative data of animals infected with AlHV-1.

Last Modified: 12/28/2014
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