Detection of EcPV-3 and EcPV-4 in paraffin-embedded aural plaque samples using PCR

• Grant number: 12/06378-2
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: June 01, 2012
• End date: May 31, 2013
• Field of knowledge: Agronomical Sciences - Veterinary Medicine - Animal Clinics and Surgery
• Principal Investigator: José Paes de Oliveira Filho
• Grantee: Luiza Stachewski Zakia
• Host Institution: Faculdade de Medicina Veterinária e Zootecnia (FMVZ). Universidade Estadual Paulista (UNESP). Campus de Botucatu. Botucatu , SP, Brazil

Abstract

Aural plaque (equine ear papilloma) is a cosmopolitan disease in distribution that causes animal discomfort and economic losses for the owners. Four types of Equus caballus papilomavirus (EcPV) were identified, and two of these were associated with aural plaque in horses, EcPV-3 and EcPV-4. DNA detection of the EcPV-3 and EcPV-4 by PCR using formalin-fixed, paraffin-embedded (FFPE) aural plaque samples has not been successfully performed. Therefore, the aim of this study will be to detect the EcPV-3 and EcPV-4 DNA by PCR, using auricular skin FFPE samples from 10 horses naturally infected. The samples were clinical and histological diagnosed as aural plaque, and EcPV-3 and/or EcPV-4 DNA were detected in all these samples (fresh samples) by PCR. DNA purification will be performed using a specially designed kit for purifying DNA from FFPE tissue sections. PCR will be performed using DNA and specific primers for EcPV-3 or EcPV-4 detection. PCR products will be purified and sequenced, and then, the sequences obtained will be compared against sequences previously deposited in GenBank, to confirm the diagnostic. The optimization of this test will allow etiologic diagnosis from aural plaque contributing to retrospective studies using archived FFPE equine aural plaque. (AU)

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

ZAKIA, L. S.; BASSO, R. M.; OLIVO, G.; HERMAN, M.; ARAUJO, JR., J. P.; BORGES, A. S.; OLIVEIRA-FILHO, J. P.. Detecção do DNA de papilomavírus em amostras de placa aural equina fixadas em formalina e embebidas em parafina. Arquivo Brasileiro de Medicina Veterinária e Zootecnia, v. 67, n. 4, p. 1193-1196, AUG 2015. (13/15995-8, 12/06378-2, 13/13973-7)