title: New PCR diagnostic systems for the detection and quantification of porcine cytomegalovirus (PCMV).
abstract: Pigs are frequently infected with porcine cytomegalovirus (PCMV). Infected adult animals may not present with symptoms of disease, and the virus remains latent. However, the virus may be transmitted to human recipients receiving pig transplants. Recently, it was shown that pig-to-non-human-primate xenotransplantations showed 2 to 3 times lower transplant survival when the donor pig was infected with PCMV. Therefore, highly sensitive methods are required to select virus-free pigs and to examine xenotransplants. Seven previously established PCR detection systems targeting the DNA polymerase gene of PCMV were examined by comparison of thermodynamic parameters of oligonucleotides, and new diagnostic nested PCR and real-time PCR systems with improved parameters and high sensitivity were established. The detection limit of conventional PCR was estimated to be 15 copies, and that of the nested PCR was 5 copies. The sensitivity of the real-time PCR with a TaqMan probe was two copies. An equal efficiency of the newly established detection systems was shown by parallel testing of DNA from sera and blood of six pigs, identifying the same animals as PCMV infected. These new diagnostic PCR systems will improve the detection of PCMV and therefore increase the safety of porcine xenotransplants.
authors.affiliation: Robert Koch Institut, Nordufer 20, 13353, Berlin, Germany. Morozovv@rki.de.
authors.forename: Vladimir A
authors.initials: VA
authors.lastname: Morozov
authors.affiliation: Engelhardt Institute of Molecular Biology RAS, Vavilova 32, 119991, Moscow, Russia.
authors.forename: Alexey V
authors.initials: AV
authors.lastname: Morozov
authors.affiliation: Robert Koch Institut, Nordufer 20, 13353, Berlin, Germany. DennerJ@rki.de.
authors.forename: Joachim
authors.initials: J
authors.lastname: Denner
mesh_terms: D000818:Animals; D003587:Cytomegalovirus / Q000235:genetics / Q000378:metabolism*; D003586:Cytomegalovirus Infections / Q000175:diagnosis / Q000662:veterinary*; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D012680:Sensitivity and Specificity; D016415:Sequence Alignment; D013552:Swine / Q000821:virology; D013553:Swine Diseases / Q000175:diagnosis* / Q000821:virology