Development of a PCR system for porcine cytomegalovirus detection and determination of the putative partial sequence of its DNA polymerase gene

• Published in: Epidemiology and infection Vol. 123; no. 1; pp. 177 - 180
• Main Authors: WIDEN, B. F., LOWINGS, J. P., BELAK, S., BANKS, M.
• Format: Journal Article
• Language: English
• Published: Cambridge Cambridge University Press 01.08.1999
• Subjects: Animal viral diseases; Animals; Antibodies; Base Sequence; Biological and medical sciences; Consensus Sequence; Cytomegalovirus; Cytomegalovirus - classification; Cytomegalovirus - genetics; Cytomegalovirus - isolation & purification; Cytomegalovirus Infections - veterinary; Cytomegalovirus Infections - virology; DNA; DNA Primers; DNA, Viral - chemistry; DNA-Directed DNA Polymerase - genetics; Fundamental and applied biological sciences. Psychology; Infections; Infectious diseases; Medical sciences; Microbiology; Molecular Sequence Data; Nucleotide sequences; Plasmids; Polymerase chain reaction; Polymerase Chain Reaction - methods; Polymerase Chain Reaction - veterinary; Porcine cytomegalovirus; Sensitivity and Specificity; SHORT PAPER; Swine; Swine Diseases - virology; Techniques used in virology; Viral diseases; Virology; Viruses; Porcine cytomegalovirus; Nucleotide sequence; Enzyme; Herpesviridae; Transferases; Method; Phylogeny; Betaherpesvirinae; Pig; Infection; Virus; Polymerase chain reaction; Nucleotidyltransferases; Vertebrata; Mammalia; Animal; Viral disease; Artiodactyla; Diagnosis; Veterinary; Detection; Ungulata; DNA-directed DNA polymerase
• DOI: 10.1017/S0950268899002599
• ISSN: 0950-2688

After PCR amplification with conservative cytomegalovirus primers, a 520 nucleotide putative partial sequence of the DNA polymerase gene of porcine cytomegalovirus (PCMV) was determined. Sequence comparison revealed homology to DNA polymerase genes from various beta herpes viruses and a dendrogram was constructed depicting the relationship of PCMV to other members of the Herpesviridae family. The dendrogram indicates that PCMV is indeed a beta herpes virus that is more closely related to human herpes virus types 6 and 7 than to type 5. To address the difficulties encountered during conventional PCMV detection and characterization a set of nested PCR primers were constructed which generated DNA fragments of 415 and 257 bp from the DNA polymerase gene. The nested PCR system proved specific for PCMV and provided a novel means for the detection of this poorly characterized herpes virus in pig populations, vaccines and in organs used in xenotransplantation.