Ultra-rapid, simple, sensitive, and economical silica method for extraction of dengue viral-RNA from clinical specimens and mosquitos 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 mul) 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-I virus was also detected in field-caught Aedes aegypti mosquitoes by silica RT-PCR.
Palavras-Chave: Dengue diagnosis; Guanidinium extraction; Genomic amplification; RNA Isolation; 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 - RNA ; Aedes aegypti - RT-PCR ; Aedes aegypti - Virus ; Aedes aegypti - Epidemic
Data de publicação: 1993
