Duplex Nested-PCR for Detection of Small Ruminant Lentiviruses

Overview

Journal Braz J Microbiol

Specialty Microbiology

Date 2018 Sep 26

PMID 30249525

Citations 11

Authors

Rebeca C Marinho

Gabrielle R Martins

Kelma C Souza

Ana Lidia M Sousa

Sabrina Tainah C Silva

Juliana A Nobre

Maria F S Teixeira

Affiliations

Soon will be listed here.

Abstract

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.

Citing Articles

Molecular Characterization of Small Ruminant Lentiviruses in Sheep and Goats: A Systematic Review.

Gobbi P, Pavone S, Orso M, Passamonti F, Righi C, Beato M Animals (Basel). 2024; 14(23).

PMID: 39682510 PMC: 11640545. DOI: 10.3390/ani14233545.

Detection and Phylogenetic Analysis of Caprine Arthritis Encephalitis Virus Using TaqMan-based qPCR in Eastern China.

Tian Y, Zhang H, Zhang Y, Zhang X, Guan Z, Zhang J Vet Sci. 2024; 11(3).

PMID: 38535872 PMC: 10975075. DOI: 10.3390/vetsci11030138.

Detection of lymphoproliferative disease virus in Iowa Wild Turkeys (Meleagris gallopavo): Comparison of two sections of the proviral genome.

Smith K, Blanchong J PLoS One. 2024; 19(2):e0296856.

PMID: 38346036 PMC: 10861079. DOI: 10.1371/journal.pone.0296856.

The genetic variability of small-ruminant lentiviruses and its impact on tropism, the development of diagnostic tests and vaccines and the effectiveness of control programmes.

Olech M J Vet Res. 2023; 67(4):479-502.

PMID: 38130459 PMC: 10730557. DOI: 10.2478/jvetres-2023-0064.

In vivo evaluation of the antiretroviral activity of Melia azedarach against small ruminant lentiviruses in goat colostrum and milk.

Souza S, Pinheiro R, Peixoto R, de Sousa A, Andrioli A, Lima A Braz J Microbiol. 2023; 55(1):875-887.

PMID: 38010582 PMC: 10920544. DOI: 10.1007/s42770-023-01174-0.

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