A multiplex DNA suspension microarray for simultaneous detection and differentiation of classical swine fever virus and other pestiviruses

Deregt, Dirk; Gilbert, Scott A.; Dudas, Sandor; Pasick, John; Baxi, Shailja; Burton, Kimberley M.; Baxi, Mohit K.

doi:10.1016/j.jviromet.2006.03.025

Cited by 46 publications

(50 citation statements)

References 18 publications

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“…Os cDNAs foram sintetizados utilizando o kit SuperScriptTM III Reverse Transcriptase (Life Technologies, USA), em volume total de 20µL. A PCR foi preparada em volume total de 25µL que continha 2µL de cDNA, usando dois protocolos para amplificar fragmentos do gene 5'NCR, com o emprego dos primers 324 5'-ATGCCCWGTAGGACTAGCA-30 (posição na cepa de BVDV-1 NADL: 108-128) e 326 5'-TCAACTCCATGTGCCATGTAC-30 (posição na cepa NADL: 395-375), que amplificam um fragmento de 288bp (Vilcek et al 1994), e 5'-CATGCCCRYAGTAGGACTAGC-30 (posição na cepa NADL: 107-127) e 5'-ATGTGCCATGTACAGCA-GAG-30 (posição na cepa NADL:: 387-368), que amplificam um fragmento de 280bp (Deregt et al 2006). Os produtos da amplificação foram marcados com Blue Green ® Loading Dye I (LGC Biotecnologia, Cotia, São Paulo, Brazil) e eletroforese em gel de agarose a 2%.…”

Section: Methodsunclassified

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Detecção do virus ‘HoBi’-like (BVDV-3) em bovino no semiárido do Estado da Paraíba

et al. 2016

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RESUMO.-Objetivou-se descrever os aspectos clínicos e anatomopatológicos, e a identificação viral de um caso de infecção pelo vírus 'Hobi'-like (BVDV-3) em bovino do semiárido paraibano, Nordeste do Brasil. Um bovino, fêmea, três meses de idade, foi levado ao Hospital Veterinário da UFCG apresentando salivação, dificuldade de apreensão do teto, falta de apetite, fezes escuras e em pouca quantidade. Diante da piora do quadro clínico optou-se por sua eutaná-sia in extremis, seguida da realização da necropsia e coleta de material para histopatologia. Histologicamente, nas mucosas do trato digestivo, havia edema, degeneração balonosa, necrose e infiltrado inflamatório, que foi observado na face dorsal da língua e no seu epitélio mais profundo. The aim of this study was to describe the clinical and anatomopathological aspects, and the viral identification of a case of 'Hobi'-like (BVDV-3) vírus infection in cattle from the semiarid of Paraiba State, Northeastern Brazil. A female bovine, three months old, was sent to the UFCG's Veterinary Hospital presenting salivation, difficulty of ceiling seizure, lack of appetite, dark feces and in small amounts. Due to worsening of symptoms it was decided to in extremis euthanasia, followed by the necropsy and collection of material for histopathology. Histologically, in the mucous membranes of the digestive tract there were edema, ballooning degeneration, necrosis and inflammatory infiltrate, which was observed on the dorsal surface of the tongue and in its deepest epithelium. The immunohistochemical of skin biopsies of the extremity of the ear (ear notches) showed positive antigenic marking and by RT-PCR it was possible to detect viral RNA of BVDV in the serum, whose cytopathic effect in epithelial cells of bovine kidney lineage "Madin Darby bovine kidney" (MDBK)was not observed. The sequencing of the 5'NCR gene showed that the virus isolated was more related to 'Hobi'-like (BVDV-3). After confirming the diagnosis serum samples were collected from 23 animals for serology by indirect ELISA, and a seropositivity of 69.6% (16/23) was found. The identification of this new case of 'Hobi'-like infection in Paraiba reaffirms the need for regular BVDV monitoring in the region to early detection of infection in the herds and adoption of effective preventive and control measures.INDEX TERMS: Cattle, BVDV-3, pestivirus, viral identification.

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Section: Methodsunclassified

“…Como controle positivo foi utilizado a cepa BVDV-1 NADL. Para o sequenciamento do DNA, sequencias parciais de 5'NCR (Deregt et al 2006), foram amplificadas por RT-PCR.…”

Section: Methodsunclassified

Detecção do virus ‘HoBi’-like (BVDV-3) em bovino no semiárido do Estado da Paraíba

et al. 2016

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“…RASOOLY AND HEROLD fluorophores (Yang et al, 2001;Summerbell et al, 2005;Deregt et al, 2006;Diaz et al, 2006;Porschewski et al, 2006;Schmitt et al, 2006). Each bead generates a unique signature (variable intensity of the two fluorophores) that provides *100 spectrally unique beads=signals that can be identified by flow cytometry.…”

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confidence: 99%

Food Microbial Pathogen Detection and Analysis Using DNA Microarray Technologies

Rasooly

Herold

2008

Foodborne Pathogens and Disease

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Culture-based methods used for microbial detection and identification are simple to use, relatively inexpensive, and sensitive. However, culture-based methods are too time-consuming for high-throughput testing and too tedious for analysis of samples with multiple organisms and provide little clinical information regarding the pathogen (e.g., antibiotic resistance genes, virulence factors, or strain subtype). DNA-based methods, such as polymerase chain reaction (PCR), overcome some these limitations since they are generally faster and can provide more information than culture-based methods. One limitation of traditional PCRbased methods is that they are normally limited to the analysis of a single pathogen, a small group of related pathogens, or a small number of relevant genes. Microarray technology enables a significant expansion of the capability of DNA-based methods in terms of the number of DNA sequences that can be analyzed simultaneously, enabling molecular identification and characterization of multiple pathogens and many genes in a single array assay. Microarray analysis of microbial pathogens has potential uses in research, food safety, medical, agricultural, regulatory, public health, and industrial settings. In this article, we describe the main technical elements of microarray technology and the application and potential use of DNA microarrays for food microbial analysis.

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“…RT-PCR assays and real-time RT-PCR, in particular, have proven to be rapid, sensitive and specific (Pfeffer et al, 1997, Scaramozzino et al, 2001, Sanchez-Seco et al, 2001, Pyke et al, 2004, Dyer et al, 2007. However, RT-PCR assays are limited in that they cannot easily be multiplexed to detect and type many viral pathogens simultaneously (Deregt et al, 2006b). For fluorescent reporting real-time RT-PCR assays, the limit for most instruments is five targets, while gel-resolved endpoint analysed molecular assays rarely effectively include more than 8 or 10 targets (Brunstein & Thomas, 2006).…”

Section: Molecular Diagnosticsmentioning

confidence: 99%

“…At the time this project commenced, the original application of the Luminex system, detection of viral nucleic acids, had only recently been applied diagnostically (Deregt et al, 2006b). The…”

Section: Suspension Microarraysmentioning

confidence: 99%

Development of multiplex suspension microarrays for the detection of alphaviruses and flaviviruses of public health significance

Fischer¹

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A multiplex DNA suspension microarray for simultaneous detection and differentiation of classical swine fever virus and other pestiviruses

Cited by 46 publications

References 18 publications

Detecção do virus ‘HoBi’-like (BVDV-3) em bovino no semiárido do Estado da Paraíba

Detecção do virus ‘HoBi’-like (BVDV-3) em bovino no semiárido do Estado da Paraíba

Food Microbial Pathogen Detection and Analysis Using DNA Microarray Technologies

Development of multiplex suspension microarrays for the detection of alphaviruses and flaviviruses of public health significance

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