Duplex nested-PCR for detection of small ruminant lentiviruses.

Autor:	Marinho RC; Universidade Estadual do Ceará, Programa de Pós-Graduação em Ciências Veterinárias, Laboratório de Virologia, Fortaleza, CE, Brazil. Electronic address: beca.marinho@hotmail.com., Martins GR; Universidade Estadual do Ceará, Programa de Pós-Graduação em Ciências Veterinárias, Laboratório de Virologia, Fortaleza, CE, Brazil., Souza KC; Embrapa Ovinos e Caprinos, Sobral, CE, Brazil., Sousa ALM; Universidade Estadual do Ceará, Programa de Pós-Graduação em Ciências Veterinárias, Laboratório de Virologia, Fortaleza, CE, Brazil., Silva STC; Universidade Estadual do Ceará, Programa de Pós-Graduação em Ciências Veterinárias, Laboratório de Virologia, Fortaleza, CE, Brazil., Nobre JA; Universidade Estadual do Ceará, Programa de Pós-Graduação em Ciências Veterinárias, Laboratório de Virologia, Fortaleza, CE, Brazil., Teixeira MFS; Universidade Estadual do Ceará, Programa de Pós-Graduação em Ciências Veterinárias, Laboratório de Virologia, Fortaleza, CE, Brazil.
Jazyk:	angličtina
Zdroj:	Brazilian journal of microbiology : [publication of the Brazilian Society for Microbiology] [Braz J Microbiol] 2018 Nov; Vol. 49 Suppl 1, pp. 83-92. Date of Electronic Publication: 2018 Aug 13.
DOI:	10.1016/j.bjm.2018.04.013
Abstrakt:	Small ruminant lentiviruses (SRLV) have high genetic variability which results in different viral strains around the world. This create a challenge to design sensible primers for molecular diagnosis in different regions. This work proposes a protocol of duplex nested-PCR for the precise diagnosis of SRLV. The technique was designed and tested with the control strains CAEV Co and MVV 1514. Then, field strains were submitted to the same protocol of duplex nested-PCR. Blood samples of sheep and goats were tested with AGID and nested PCR with specific primers for pol, gag and LTR. The AGID results showed low detection capacity of positive animals, while the nested PCR demonstrated a greater capacity of virus detection. Results demonstrated that LTR-PCR was more efficient in detecting positive sheep samples, whereas gag-PCR allowed a good detection of samples of positive goats and positive sheep. In addition, pol-PCR was more efficient with goat samples than for sheep. Duplex nested PCR performed with standard virus samples and field strains demonstrated that the technique is more efficient for the detection of multiple pro-viral DNA sequences. This study demonstrated a successful duplex nested PCR assay allowing a more accurate diagnosis of SRLV. (Copyright © 2018 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.)
Databáze:	MEDLINE
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