title: Bovine papillomavirus isolation by ultracentrifugation.
abstract: The bovine papillomavirus (BPV) is the etiological agent of bovine papillomatosis, which causes significant economic losses to livestock, characterized by the presence of papillomas that regress spontaneously or persist and progress to malignancy. Currently, there are 13 types of BPVs described in the literature as well as 32 putative new types. This study aimed to isolate viral particles of BPV from skin papillomas, using a novel viral isolation method. The virus types were previously identified with new primers designed. 77 cutaneous papilloma samples of 27 animals, Simmental breed, were surgically removed. The DNA was extracted and subjected to PCR using Delta-Epsilon and Xi primers. The bands were purified and sequenced. The sequences were analyzed using software and compared to the GenBank database, by BLAST tool. The viral typing showed a prevalence of BPV-2 in 81.81% of samples. It was also detected the presence of the putative new virus type BR/UEL2 in one sample. Virus isolation was performed by ultracentrifugation in a single density of cesium chloride. The method of virus isolation is less laborious than those previously described, allowing the isolation of complete virus particles of BPV-2.
authors.affiliation: Laboratório de Genética, Instituto Butantan, Av. Vital Brasil, 1500, São Paulo 05503-900, SP, Brazil; Programa de Pós-graduação Interunidades em Biotecnologia, Universidade de São Paulo, Av. Prof. Lineu Prestes, 2415, Ed. ICB III, Cidade Universitária, São Paulo 05508-900, SP, Brazil. Electronic address: rodrigo.araldi@butantan.gov.br.
authors.forename: R P
authors.initials: RP
authors.lastname: Araldi
authors.affiliation: Programa de Pós-graduação Interunidades em Biotecnologia, Universidade de São Paulo, Av. Prof. Lineu Prestes, 2415, Ed. ICB III, Cidade Universitária, São Paulo 05508-900, SP, Brazil; Laboratório de Parasitologia, Instituto Butantan, Av. Vital Brasil, 1500, São Paulo 05503-900, SP, Brazil. Electronic address: dalton.silva@butantan.gov.br.
authors.forename: D N S
authors.initials: DN
authors.lastname: Giovanni
authors.affiliation: Laboratório de Genética, Instituto Butantan, Av. Vital Brasil, 1500, São Paulo 05503-900, SP, Brazil; Programa de Pós-graduação em Biologia Estrutural e Funcional, Universidade Federal de São Paulo, Ed. Leitão da Cunha, R. Botucatu, 740, São Paulo 04023-900, SP, Brazil. Electronic address: thatianacmelo@yahoo.com.br.
authors.forename: T C
authors.initials: TC
authors.lastname: Melo
authors.affiliation: Laboratório de Genética, Instituto Butantan, Av. Vital Brasil, 1500, São Paulo 05503-900, SP, Brazil. Electronic address: nara.diniz@gmail.com.
authors.forename: N
authors.initials: N
authors.lastname: Diniz
authors.affiliation: Laboratório de Genética, Instituto Butantan, Av. Vital Brasil, 1500, São Paulo 05503-900, SP, Brazil; Programa de Pós-graduação Interunidades em Biotecnologia, Universidade de São Paulo, Av. Prof. Lineu Prestes, 2415, Ed. ICB III, Cidade Universitária, São Paulo 05508-900, SP, Brazil. Electronic address: jackmazzuchelli@yahoo.com.br.
authors.forename: J
authors.initials: J
authors.lastname: Mazzuchelli-de-Souza
authors.affiliation: Laboratório de Genética, Instituto Butantan, Av. Vital Brasil, 1500, São Paulo 05503-900, SP, Brazil; Programa de Pós-graduação Interunidades em Biotecnologia, Universidade de São Paulo, Av. Prof. Lineu Prestes, 2415, Ed. ICB III, Cidade Universitária, São Paulo 05508-900, SP, Brazil. Electronic address: thalita.a.santana@hotmail.com.
authors.forename: T A
authors.initials: TA
authors.lastname: Sant'Ana
authors.affiliation: Laboratório de Genética, Instituto Butantan, Av. Vital Brasil, 1500, São Paulo 05503-900, SP, Brazil. Electronic address: rodsfc@butantan.gov.br.
authors.forename: R F
authors.initials: RF
authors.lastname: Carvalho
authors.affiliation: Laboratório de Genética, Instituto Butantan, Av. Vital Brasil, 1500, São Paulo 05503-900, SP, Brazil; Departamento de Biologia, Universidade Federal da Integração Latino-Americana, Av. Silvio Américo Sasdelli, 1842, Vila A, Ed. Comercial Lorivo, Foz do Iguaçú 85866-000, PR, Brazil. Electronic address: wbecak@pq.cnpq.br.
authors.forename: W
authors.initials: W
authors.lastname: Beçak
authors.affiliation: Laboratório de Genética, Instituto Butantan, Av. Vital Brasil, 1500, São Paulo 05503-900, SP, Brazil; Programa de Pós-graduação Interunidades em Biotecnologia, Universidade de São Paulo, Av. Prof. Lineu Prestes, 2415, Ed. ICB III, Cidade Universitária, São Paulo 05508-900, SP, Brazil. Electronic address: rita.stocco@butantan.gov.br.
authors.forename: R C
authors.initials: RC
authors.lastname: Stocco
mesh_terms: D000818:Animals; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis* / Q000821:virology; D004279:DNA, Viral / Q000737:chemistry / Q000235:genetics; D027383:Papillomaviridae / Q000235:genetics / Q000302:isolation & purification*; D030361:Papillomavirus Infections / Q000662:veterinary* / Q000821:virology; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D012867:Skin / Q000821:virology; D014461:Ultracentrifugation / Q000379:methods*; D014771:Virion / Q000235:genetics / Q000302:isolation & purification; D014773:Virology / Q000379:methods*
chemical_list: D004279:DNA, Viral
keywords: Bovine papillomavirus; Diagnosis; Ultracentrifugation; Virus isolation