Araldi R P, Giovanni D N S, Melo T C, Diniz N, Mazzuchelli-de-Souza J, Sant'Ana T A, Carvalho R F, Beçak W, Stocco R C
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.
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.
J Virol Methods. 2014 Nov;208:119-24. doi: 10.1016/j.jviromet.2014.07.029. Epub 2014 Aug 4.
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.
牛乳头瘤病毒（BPV）是牛乳头瘤病的病原体，会给家畜造成重大经济损失，其特征是出现可自发消退或持续存在并发展为恶性肿瘤的乳头瘤。目前，文献中描述了13种BPV类型以及32种假定的新类型。本研究旨在使用一种新型病毒分离方法从皮肤乳头瘤中分离BPV病毒颗粒。此前已用设计的新引物鉴定了病毒类型。手术切除了27头西门塔尔牛品种动物的77个皮肤乳头瘤样本。提取DNA并使用Delta-Epsilon和Xi引物进行PCR。对条带进行纯化和测序。使用软件对序列进行分析，并通过BLAST工具与GenBank数据库进行比较。病毒分型显示81.81%的样本中BPV-2占优势。在一个样本中还检测到假定的新病毒类型BR/UEL2的存在。通过在单一密度的氯化铯中进行超速离心来进行病毒分离。该病毒分离方法比先前描述的方法省力，能够分离出BPV-2的完整病毒颗粒。
