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Your search keyword '"Ferreira-Machado E"' showing total 2 results
Start Over Author "Ferreira-Machado E" Topic real-time polymerase chain reaction
2 results on '"Ferreira-Machado E"'

1. Novel and highly sensitive SYBR® Green real-time pcr for poxvirus detection in odontocete cetaceans.

Author

Sacristán C, Catão-Dias JL, Ewbank AC, Ferreira-Machado E, Neves E, Santos-Neto EB, Azevedo A, Laison-Brito J Jr, De Castilho PV, Daura-Jorge FG, Simões-Lopes PC, Carballo M, García-Párraga D, Sánchez-Vizcaíno JM, and Esperón F

Subjects
Animals, Benzothiazoles, DNA Primers genetics, Diamines, Poxviridae genetics, Poxviridae Infections diagnosis, Quinolines, Sensitivity and Specificity, Cetacea virology, Molecular Diagnostic Techniques methods, Organic Chemicals metabolism, Poxviridae isolation & purification, Poxviridae Infections veterinary, Real-Time Polymerase Chain Reaction methods, Staining and Labeling methods
Abstract

Poxviruses are emerging pathogens in cetaceans, temporarily named 'Cetaceanpoxvirus' (CePV, family Poxviridae), classified into two main lineages: CePV-1 in odontocetes and CePV-2 in mysticetes. Only a few studies performed the molecular detection of CePVs, based on DNA-polymerase gene and/or DNA-topoisomerase I gene amplification. Herein we describe a new real-time PCR assay based on SYBR ® Green and a new primer set to detect a 150 bp fragment of CePV DNA-polymerase gene, also effective for conventional PCR detection. The novel real-time PCR was able to detect 5 up to 5 × 10 6 copies per reaction of a cloned positive control. Both novel PCR methods were 1000 to 100,000-fold more sensitive than those previously described in the literature. Samples of characteristic poxvirus skin lesions ('tattoo') from one Risso's dolphin (Grampus griseus), two striped dolphins (Stenella coeruleoalba) and two Guiana dolphins (Sotalia guianensis) were all positive to both our novel real time- and conventional PCR methods, even though three of these animals (a Risso's dolphin, a striped dolphin, and a Guiana dolphin) were previously negative to the conventional PCRs previously available. To our knowledge, this is the first real-time PCR detection method for Cetaceanpoxvirus, a much more sensitive tool for the detection of CePV-1 infections., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Published
2018
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2. Novel and highly sensitive SYBR® Green real-time PCR for poxvirus detection in odontocete cetaceans

Author

Sacristán, C., Catão-Dias, J. L., Ewbank, A. C., Ferreira-Machado, E., Neves, E., Santos-Neto, E. B., Azevedo, A., Laison-Brito, J., Jr., De Castilho, P. V., Daura-Jorge, F. G., Simões-Lopes, P. C., Carballo, M., García-Párraga, D., Sánchez-Vizcaíno, J. M., and Esperón, F.

Subjects
0301 basic medicine, Tattoo skin lesion, Sotalia guianensis, 030106 microbiology, Poxviridae Infections, Stenella coeruleoalba, Diamines, Real-Time Polymerase Chain Reaction, Sensitivity and Specificity, law.invention, Brazil Cetacean, 03 medical and health sciences, law, Virology, biology.animal, Gene duplication, Animals, Benzothiazoles, Grampus griseus, Organic Chemicals, Gene, Polymerase chain reaction, DNA Primers, Staining and Labeling, biology, Poxviridae, biology.organism_classification, 030104 developmental biology, Real-time polymerase chain reaction, Molecular Diagnostic Techniques, Poxvirus, Mediterranean sea, Quinolines, Cetacea, Primer (molecular biology), REAÇÃO EM CADEIA POR POLIMERASE, human activities
Abstract

Poxviruses are emerging pathogens in cetaceans, temporarily named ‘Cetaceanpoxvirus’ (CePV, family Poxviridae), classified into two main lineages: CePV-1 in odontocetes and CePV-2 in mysticetes. Only a few studies performed the molecular detection of CePVs, based on DNA-polymerase gene and/or DNA-topoisomerase I gene amplification. Herein we describe a new real-time PCR assay based on SYBR® Green and a new primer set to detect a 150 bp fragment of CePV DNA-polymerase gene, also effective for conventional PCR detection. The novel real-time PCR was able to detect 5 up to 5 × 106 copies per reaction of a cloned positive control. Both novel PCR methods were 1000 to 100,000-fold more sensitive than those previously described in the literature. Samples of characteristic poxvirus skin lesions (‘tattoo’) from one Risso's dolphin (Grampus griseus), two striped dolphins (Stenella coeruleoalba) and two Guiana dolphins (Sotalia guianensis) were all positive to both our novel real time- and conventional PCR methods, even though three of these animals (a Risso's dolphin, a striped dolphin, and a Guiana dolphin) were previously negative to the conventional PCRs previously available. To our knowledge, this is the first real-time PCR detection method for Cetaceanpoxvirus, a much more sensitive tool for the detection of CePV-1 infections.

Published
2018

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