title: Application of conventional and real-time fluorescent ITS1 rDNA PCR for detection of Besnoitia besnoiti infections in bovine skin biopsies.
abstract: Besnoitia besnoiti, an apicomplexan protozoan parasite, is the causative agent of bovine besnoitiosis. This infection may dramatically affect body condition, and, in males, lead to irreversible infertility. While identification of clinical cases and their histopathological confirmation is relatively simple to carry out, finding subclinical forms of infection is more difficult, thus a more sensitive test for the identification of the etiological agent may be an appropriate diagnostic tool. We have developed the ITS1 rDNA-sequence-based conventional and real-time PCR which are highly sensitive and specific for the detection of B. besnoiti infection in cattle. A recombinant internal positive control was introduced to assess possible sample-related inhibitory effects during the amplification reaction and, in order to prevent false-positive results, a pre-PCR treatment of potentially contaminating dU-containing PCR product with uracil-DNA-glycosylase (UDG) was followed.
authors.affiliation: Laboratório de Parasitologia, Núcleo da Mitra, ICAM, Universidade de Evora, Apartado 94, 7000-554 Evora, Portugal. hcec@uevora.pt
authors.forename: Helder C E
authors.initials: HC
authors.lastname: Cortes
authors.forename: Yara
authors.initials: Y
authors.lastname: Reis
authors.forename: Bruno
authors.initials: B
authors.lastname: Gottstein
authors.forename: Andrew
authors.initials: A
authors.lastname: Hemphill
authors.forename: Alexandre
authors.initials: A
authors.lastname: Leitão
authors.forename: Norbert
authors.initials: N
authors.lastname: Müller
mesh_terms: D000818:Animals; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis / Q000469:parasitology*; D003048:Coccidiosis / Q000175:diagnosis / Q000469:parasitology / Q000662:veterinary*; D021903:DNA, Ribosomal Spacer / Q000032:analysis / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000662:veterinary*; D021745:Sarcocystidae / Q000235:genetics* / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D012867:Skin / Q000469:parasitology*
chemical_list: D021903:DNA, Ribosomal Spacer