Improvement of a RT-PCR assay for Yellow Fever virus genome detection
Rev. ciênc. farm. básica apl
;
38(1)2017. ilus
Article
in English
| LILACS
| ID: biblio-964195
ABSTRACT
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 diï¬erent vectors and hosts is required to avoid negative impacts on human health, tourism and trade.(AU)
Full text:
Available
Index:
LILACS (Americas)
Main subject:
Yellow Fever
/
Reverse Transcriptase Polymerase Chain Reaction
/
Flavivirus
Type of study:
Diagnostic study
Limits:
Animals
/
Humans
Language:
English
Journal:
Rev. ciênc. farm. básica apl
Journal subject:
Pharmacology
Year:
2017
Type:
Article
Affiliation country:
Brazil
Institution/Affiliation country:
Instituto Evandro Chagas/BR
/
Universidade Federal da Integração Latino-Americana/BR
/
Universidade Federal do Paraná (UFPR)/BR
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