Ultra-rapid, simple, sensitive, and economical silica method for extraction of dengue viral RNA from clinical specimens and mosquitoes by reverse transcriptase-polymerase chain reaction
Autor(es): Chungue E,Roche C,Lefevre M F,Barbazan P,Chanteau S
Resumo: A rapid, simple and efficient single-tube procedure is described for the isolation of dengue virus RNA from small amount of serum (10 microliters) followed by a reverse transcriptase-polymerase chain reaction (RT-PCR). Recovery of RNA is based on the lysing and nuclease-inactivating properties of guanidinium thiocyanate in the presence of silica. The silica RT-PCR can be completed within 5 hours starting from RNA extraction to agarose gel electrophoresis. All of the 63 dengue-3 culture-positive sera were RT-PCR-positive (virus titres: < 10(2) to 11(10.69.). Of 33 culture-negative acute sera from serologically confirmed dengue fever patients collected during dengue-3 epidemic, 4 were RT-PCR-positive. RT-PCR was also positive in 29 of 30 dengue-1 culture-positive sera (virus titres range: < 10(2) to 10(8.69). Dengue-1 virus was also detected in field-caught Aedes aegypti mosquitoes by silica RT-PCR.
Imprenta: Journal of Medical Virology, v. 40, n. 2, p. 142-145, 1993
Identificador do objeto digital: 10.1002/jmv.1890400211
Descritores: Aedes aegypti - DNA ; Aedes aegypti - Flaviviridae ; Aedes aegypti - Pathogenesis ; Aedes aegypti - Proteins ; Aedes aegypti - RNA ; Aedes aegypti - Viral infections ; Aedes aegypti - RT-PCR ; Aedes aegypti - virus ; Aedes aegypti - Dengue ; Aedes aegypti - Epidemic ; Aedes aegypti - Public health
Data de publicação: 1993
