Open AccessDiscrimination between sheep-associated and wildebeest-associated malignant catarrhal fever virus by means of a single-tube duplex nested PCR
Author(s) -
C.W. Bremer,
H. Swart,
Fulufhelo Amanda Doboro,
Baptiste Dungu,
Marco Romito,
G. J. Viljoen
Publication year - 2005
Publication title -
onderstepoort journal of veterinary research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.563
H-Index - 35
eISSN - 2219-0635
pISSN - 0030-2465
DOI - 10.4102/ojvr.v72i4.184
Subject(s) - amplicon , nested polymerase chain reaction , virology , biology , polymerase chain reaction , primer (cosmetics) , recombinase polymerase amplification , duplex (building) , gene , virus , dna , microbiology and biotechnology , genetics , chemistry , organic chemistry
A single-tube duplex nested polymerase chain reaction (sdn-PCR) was developed for the detection of and discrimination between ovine herpesvirus-2 (OvHV-2) and alcelaphine herpesvirus-1 (AIHV-1). These viruses respectively cause sheep- and wildebeest-associated malignant catarrhal fever (SA-MCF and WA-MCF). In the first step of the sdn-PCR, two primers with high annealing temperatures based on conserved regions of the tegument genes were used for DNA amplification. In the second step, two primer sets based on variable regions of the respective OvHV-2 and AIHV-1 genes and with annealing temperatures > 11 degrees C below the primers used in the first step, were used. Internal regions of different sizes from amplicons produced in the first step were amplified. This single-tube test obviates the need for two separate assays to detect both viral types, thereby reducing time, labour and cost.