Improvement of a RT-PCR assay for Yellow Fever virus genome detection
The aim of the present study was to describe an improved protocol of reverse transcription polymerase chain reaction (RT-PCR) for Yellow Fever virus genome detection. A strain of ribonucleic acid of Yellow Fever virus was submitted to the improved protocol of RT-PCR and the amplicons were visualized...
| Autor principal: | Rocha, Tatiana Carneiro da |
|---|---|
| Outros Autores: | Silva, Mario Antonio Navarro, Bona, Ana Caroline Dalla, Nunes, M?rcio Roberto Teixeira, Svoboda, Walfrido Huhl, Gomes, Eliane Carneiro |
| Grau: | Artigo |
| Idioma: | eng |
| Publicado em: |
School of Pharmaceutical Sciences at the S?o Paulo State University
2019
|
| Assuntos: | |
| Acesso em linha: |
http://patua.iec.gov.br//handle/iec/3637 |
| id |
ir-iec-3637 |
|---|---|
| recordtype |
dspace |
| spelling |
ir-iec-36372019-04-11T19:16:53Z Improvement of a RT-PCR assay for Yellow Fever virus genome detection Aperfei?oamento de um ensaio de RT-PCR para detec??o do genoma do v?rus da Febre Amarela Rocha, Tatiana Carneiro da Silva, Mario Antonio Navarro Bona, Ana Caroline Dalla Nunes, M?rcio Roberto Teixeira Svoboda, Walfrido Huhl Gomes, Eliane Carneiro Febre Amarela / gen?tica Rea??o em Cadeia da Polimerase / m?todos Rea??o em Cadeia da Polimerase em Tempo Real / m?todos Flavivirus / patogenicidade Genoma Viral / gen?tica The aim of the present study was to describe an improved protocol of reverse transcription polymerase chain reaction (RT-PCR) for Yellow Fever virus genome detection. A strain of ribonucleic acid of Yellow Fever virus was submitted to the improved protocol of RT-PCR and the amplicons were visualized under ultraviolet transilluminator, purifed and sequenced. The nucleotide sequence obtained was compared with sequences available in GenBank using the tblastx tool. The amplicons produced by the strain of ribonucleic acid of Yellow Fever virus exhibited fragments of 400 and 800 base pairs and the consensus sequence exhibited a similarity of 100% with Yellow Fever virus sequences recorded in GenBank. The improved protocol described in this study allowed Yellow Fever virus genome detection and enabled the elimination of the nested-PCR step, which has been frequently associated with contamination. In addition, it reduced the time of reaction, the cost of reagents and the possibility of sample contamination. New methods of investigating these infections must be elaborated and a continuous vigilance of these viruses in their di?erent vectors and hosts is required to avoid negative impacts on human health, tourism and trade. Coordination of Training of Higher Education Graduate Foundation [Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior (CAPES)] for providing the financial support and Funda??o Arauc?ria, Pharmaceutical Sciences graduate program ? Universidade Federal do Paran?. 2019-04-11T18:00:21Z 2019-04-11T18:00:21Z 2017 Artigo ROCHA, Tatiana Carneiro da et al. Improvement of a RT-PCR assay for Yellow Fever virus genome detection. Revista de Ci?ncias Farmac?uticas B?sica e Aplicada, v. 38, n. 1, p. 1-4, 2017. 2179-443X http://patua.iec.gov.br//handle/iec/3637 eng Acesso Aberto application/pdf School of Pharmaceutical Sciences at the S?o Paulo State University |
| institution |
Instituto Evandro Chagas (IEC) |
| collection |
PATUA |
| language |
eng |
| topic |
Febre Amarela / gen?tica Rea??o em Cadeia da Polimerase / m?todos Rea??o em Cadeia da Polimerase em Tempo Real / m?todos Flavivirus / patogenicidade Genoma Viral / gen?tica |
| spellingShingle |
Febre Amarela / gen?tica Rea??o em Cadeia da Polimerase / m?todos Rea??o em Cadeia da Polimerase em Tempo Real / m?todos Flavivirus / patogenicidade Genoma Viral / gen?tica Rocha, Tatiana Carneiro da Improvement of a RT-PCR assay for Yellow Fever virus genome detection |
| topic_facet |
Febre Amarela / gen?tica Rea??o em Cadeia da Polimerase / m?todos Rea??o em Cadeia da Polimerase em Tempo Real / m?todos Flavivirus / patogenicidade Genoma Viral / gen?tica |
| description |
The aim of the present study was to describe an improved protocol of reverse transcription polymerase chain reaction (RT-PCR) for Yellow Fever virus genome detection. A strain of ribonucleic acid of Yellow Fever virus was submitted to the improved protocol of RT-PCR and the amplicons were visualized under ultraviolet transilluminator, purifed and sequenced. The nucleotide sequence obtained was compared with sequences available in GenBank using the tblastx tool. The amplicons produced by the strain of ribonucleic acid of Yellow Fever virus exhibited fragments of 400 and 800 base pairs and the consensus sequence exhibited a similarity of 100% with Yellow Fever virus sequences recorded in GenBank. The improved protocol described in this study allowed Yellow Fever virus genome detection and enabled the elimination of the nested-PCR step, which has been frequently associated with contamination. In addition, it reduced the time of reaction, the cost of reagents and the possibility of sample contamination. New methods of investigating these infections must be elaborated and a continuous vigilance of these viruses in their di?erent vectors and hosts is required to avoid negative impacts on human health, tourism and trade. |
| format |
Artigo |
| author |
Rocha, Tatiana Carneiro da |
| author2 |
Silva, Mario Antonio Navarro Bona, Ana Caroline Dalla Nunes, M?rcio Roberto Teixeira Svoboda, Walfrido Huhl Gomes, Eliane Carneiro |
| author2Str |
Silva, Mario Antonio Navarro Bona, Ana Caroline Dalla Nunes, M?rcio Roberto Teixeira Svoboda, Walfrido Huhl Gomes, Eliane Carneiro |
| title |
Improvement of a RT-PCR assay for Yellow Fever virus genome detection |
| title_short |
Improvement of a RT-PCR assay for Yellow Fever virus genome detection |
| title_full |
Improvement of a RT-PCR assay for Yellow Fever virus genome detection |
| title_fullStr |
Improvement of a RT-PCR assay for Yellow Fever virus genome detection |
| title_full_unstemmed |
Improvement of a RT-PCR assay for Yellow Fever virus genome detection |
| title_sort |
improvement of a rt-pcr assay for yellow fever virus genome detection |
| publisher |
School of Pharmaceutical Sciences at the S?o Paulo State University |
| publishDate |
2019 |
| url |
http://patua.iec.gov.br//handle/iec/3637 |
| _version_ |
1717584476959145984 |
| score |
11.653393 |