Quantification of ovine herpesvirus 2 by digital PCR in an outbreak of malignant catarrhal fever

Oliveira, Tatiana Flávia Pinheiro de; Laguardia-Nascimento, Mateus; Xavier, Fabiana Galtarossa; Pinto, Carla do Amaral; Ferreira, Luciana Rabello; Souza, Ivy de Castro Campos de; Hammerschmitt, Márcia Elisa; Bianchi, Ronaldo Michel; Wronski, Júlia Gabriela; Etges, Rodrigo Nestor; Rigon, Grazziane Maciel; Camargos, Marcelo Fernandes; Júnior, Anselmo Vasconcelos Rivetti; Fonseca, Antônio Augusto

doi:10.1007/s00705-019-04382-y

Cited by 8 publications

(4 citation statements)

References 18 publications

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“…De-Falco et al used ddPCR to detect BPV (bovine papillomavirus) DNA in blood samples for cows, and the positivity rate was significantly higher than that of qPCR [ 29 ]. de Oliveira et al reported that ddPCR was able to detect low copy numbers of OvHV-2 in sheep and cattle [ 30 ]. de Oliveira et al developed a novel RT-ddPCR method for the detection of FMDV RNA, and it was used to test 60 bovine samples from animals with suspected vesicular diseases that were previously tested by RT-qPCR [ 31 ].…”

Section: Discussionmentioning

confidence: 99%

Development of a droplet digital PCR assay to detect bovine alphaherpesvirus 1 in bovine semen

Zhao

Chen

et al. 2022

BMC Vet Res

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Background Infectious bovine rhinotracheitis (IBR) caused by bovine alphaherpesvirus 1 (BoHV-1) is one of the most important contagious diseases in bovine. This is one of the most common infectious disease of cattle. This has led to high economic losses in the cattle farming industry. BoHV-1 can potentially be transmitted via semen during natural or artificial insemination (AI). Therefore, testing methods for the early diagnosis of BoHV-1 infection are urgently needed for international trade of ruminant semen. In this study, we developed a novel droplet digital PCR (ddPCR) assay for the detection of BoHV-1 DNA in semen samples. Results The ddPCR results showed that the detection limit was 4.45 copies per reaction with high reproducibility. The established method was highly specific for BoHV-1 and did not show cross-reactivity with specify the organisms (BTV, BVDV, Brucella, M . bovis). The results of clinical sample testing showed that the positivity rate of ddPCR (87.8%) was higher than that of qPCR (84.1%). Conclusions The ddPCR assay showed good accuracy for mixed samples and could be a new added diagnostic tool for detecting BoHV-1.

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Section: Discussionmentioning

confidence: 99%

Development of a droplet digital PCR assay to detect bovine alphaherpesvirus 1 in bovine semen

Zhao

Chen

et al. 2022

BMC Vet Res

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“…A infecção com Gammaherpesvirus 2 ovino (OvHV-2) é geralmente assintomática em ovelhas, no entanto, ocorre spillover (infecção por transbordamento, transmissão) entre espécies, causando a enfermidade em bovinos (PINHEIRO DE OLIVEIRA et al, 2019).…”

Section: Discussionunclassified

Ocorrência De Orthopoxvirus Em Animais No Brasil: Revisão De Literatura

Arruda¹,

Coimbra²,

Bezerra³

et al. 2021

Epidemiologia: Estudos Clínicos E Revisões Bibliográficas

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A obra intitulada: "EPIDEMIOLOGIA: ESTUDOS CLÍNICOS E REVISÕES" reflete sobre a Epidemiologia e a interface com Atenção Primária a Saúde, Infecções Relacionadas à Assistência à Saúde (IRAS), Pandemia provocada pela COVID-19, Oncologia, entre outros. Nesse sentido, faz-se necessário compreender a epidemiologia como um ramo da ciência que estuda o processo saúdedoença e contribui com a construção de políticas públicas direcionadas para o controle dos problemas e agravos a saúde. Em nossos livros selecionamos um dos capítulos para premiação como forma de incentivo para os autores, e entre os excelentes trabalhos selecionados para compor este livro, o premiado foi o capítulo 17, intitulado "PRÁTICA E MOTIVOS ATRIBUÍDOS AO USO DE MÁSCARAS ENTRE ESTUDANTES DA SAÚDE DURANTE A PANDEMIA DA COVID-19".Neste ínterim, destaca-se que diante do cenário atual de saúde pública provocado pela COVID-19 identificar os fatores motivadores para a prática do uso de máscaras é fundamental para auxiliar no desenvolvimento de ações de incentivo a esse cuidado essencial para o enfrentamento da pandemia. Assim, espera-se enriquecer a produção científica sobre epidemiologia, agregar o conhecimento científico, subsidiar conhecimento dos profissionais, estudantes e sociedade para compreensão do cenário de saúde atual, e possibilitar reflexões que possam incentivar outros estudos para fortalecer a pesquisa no Brasil pautadas nas evidências científicas.

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“…Samples were eluted in a final volume of 30 μL of DNase/RNase-free water. RT-qPCR was performed for the 18S gene to verify RNA extraction [ 13 ] (PINHEIRO DE OLIVEIRA et al, 2019).…”

Section: Methodsmentioning

confidence: 99%

“…To determine the annealing temperature of each pair of primers for digital droplet PCR (ddPCR), temperature gradients ranging from 54 to 64 °C were performed. The choice of temperature was based on the number of droplets above 10,000 and the greater amplitude of separation of positive and negative drops [ 13 ] (Pinheiro-de-Oliveira et al, 2018). Thus, an annealing temperature of 58 °C for primers VSAV.GP.95, 60 °C for VSAV.GP.78, and 54 °C for VSAV.PPP.77 was settled.…”

Section: Methodsmentioning

confidence: 99%

Quantification of in vitro replication kinetics of Alagoas vesiculovirus isolates by digital droplet RT-PCR

et al. 2023

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Vesicular stomatitis caused by Alagoas vesiculovirus (VSAV) has generated disease outbreaks in Brazil, mainly in the northeast region. Phylogenetic studies divide the isolates into three distinct genotypes (A, B, and C). However, there is no description of how this genetic divergence reflects on the phenotype of VSAV isolates such as in vitro replication fitness. Therefore, the objective of this work was to evaluate the ability of three distinct genotypes of Brazilian isolates of VSAV to grow in different cell-culture lines (BHK-21, Vero, and NCI-H1299). Quantification of viral RNA was performed using RT-PCR digital droplet from supernatant of cell culture collected every 4 h for a period of 24 h of viral growth in three different cell lines (BHK-21, Vero, and NCI-H1299). It was observed that the genotype C isolate has the lowest replication efficiency among the three analyzed viruses, without major changes in the copies of viral RNA over the entire time of the study.

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Quantification of ovine herpesvirus 2 by digital PCR in an outbreak of malignant catarrhal fever

Cited by 8 publications

References 18 publications

Development of a droplet digital PCR assay to detect bovine alphaherpesvirus 1 in bovine semen

Development of a droplet digital PCR assay to detect bovine alphaherpesvirus 1 in bovine semen

Ocorrência De Orthopoxvirus Em Animais No Brasil: Revisão De Literatura

Quantification of in vitro replication kinetics of Alagoas vesiculovirus isolates by digital droplet RT-PCR

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