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United States Department of Agriculture

Agricultural Research Service

Title: Pcr Detection of North American and Central African Isolates of Epixootic Hemorrhagic Disease Virus (Ehdv) Based on Genome Segment 10 of Ehdv Serotype 1

item Wilson, William
item Schore, C - UNIV CA - DAVIS
item Mohammed, M. E. - UNIV CA - DAVIS
item Yilma, T - UNIV CA - DAVIS
item Cullor, J - UNIV CA - DAVIS
item Osburn, B - UNIV CA - DAVIS

Submitted to: Journal of Clinical Microbiology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: June 12, 1998
Publication Date: September 1, 1998
Citation: Aradaib, I.E., Wilson, W.C., Schore, C.E., Mohammed, M.H., Yilma, T.D., Cullor, J.S., Osburn, B.I., 1998. Pcr detection of north american and central african isolates of epixootic hemorrhagic disease virus (ehdv) based on genome segment 10 of ehdv serotype 1. Journal of Clinical Microbiology. 36:2604-2608.

Interpretive Summary: This paper compares describes the development of procedures to detect epizootic hemorrhagic disease viral RNA in clinical specimens using a gene amplification test.

Technical Abstract: PCR amplification technology for the detection of epizootic hemorrhagic disease virus (EHDV) ribonucleic acid in cell culture and clinical specimens was developed. With oligoribonucleotide primers selected from genome segment 10 of EHDV serotype 1 (EHDV-1), which codes for two nonstructural proteins (NS3 and NS3a), the PCR-based assay resulted in a 535-bp PCR product. RNAs from North American EHDV-1 prototype, EHDV-2 prototype, and a number of EHDV field isolates, including the Central African isolates of EHDV-5 and EHDV-318 propagated in cell cultures, were detected by this PCR-based assay. The specific 535-bp PCR products were visualized onto agarose gels, and the identity of the PCR products was confirmed by chemiluminescent hybridization with a 352-bp internal probe. The sensitivity of the EHDV PCR assay was increased by chemiluminescent hybridization; by this EHDV-NS3 PCR, 10 fg of EHDV RNA was detected (equivalent to 600 viral particles). Amplification product was not detected when the PCR-based assay was applied to RNAs from North American bluetongue virus prototype serotypes 2, 10, 11, 13, and 17; total nucleic acid extracts from uninfected BHK-21 cells; or unfractionated blood from calves and deer that were EHDV seronegative and virus isolation negative. The described EHDV PCR-based assay with primers derived from segment 10 of EHDV-1 resulted in detection of EHDV RNA from blood and tissues collected from calves and deer with natural and experimental EHDV infections and provides a valuable tool to study the epidemiology of EHDV infection in susceptible ruminants. Please reference the following: (1) HQ issued Clearance/Sensitivity Responsibility memo (attached), and also found by clicking on the Help button when in the ARIS Project Info tab for ARS-115 (2) NPA Policy Memorandum PM-03-006, dated 06/25/2003 Subject: Submission of the ARS-115/Publication in ARIS, Northern Plains Area

Last Modified: 4/22/2015
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