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 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 (approximately with 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.
Traul
Donald L
DL
Animal Diseases Research Unit, Agricultural Research Service, United States Department of Agriculture, Washington State University, Pullman, 99164, USA. dtraul@vetmed.wsu.edu
Elias
Shirley
S
Taus
Naomi S
NS
Herrmann
Lynn M
LM
Oaks
J Lindsay
JL
Li
Hong
H
D000818:Animals; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis* / Q000821:virology; D018142:Gammaherpesvirinae / Q000235:genetics / Q000302:isolation & purification*; D006566:Herpesviridae Infections / Q000175:diagnosis / Q000662:veterinary*; D016133:Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D014412:Tumor Virus Infections / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D015678:Viral Structural Proteins / Q000235:genetics
D015678:Viral Structural Proteins