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

A rapid, simple and efficient single‐tube procedure is described for the isolation of dengue virus RNA from small amount of serum (10 μl) followed by a reverse transcriptase‐polymerase chain reaction (RT‐PCR). Recovery of RNA is based on the lysing and nuclease‐inactivating properties of guanidinium...

Ausführliche Beschreibung

Gespeichert in:

Bibliographische Detailangaben
Veröffentlicht in:	Journal of medical virology 1993-06, Vol.40 (2), p.142-145
Hauptverfasser:	Chungue, Eliane, Roche, Claudine, Lefevre, Marie-France, Barbazan, P., Chanteau, Suzanne
Format:	Artikel
Sprache:	eng
Schlagworte:	Aedes - microbiology Aedes aegypti Animals Arboviroses Biological and medical sciences Dengue - microbiology dengue diagnosis Dengue fevers dengue virus Dengue Virus - genetics Dengue Virus - isolation & purification Female genomic amplification Guanidines guanidinium extraction Human viral diseases Humans Infectious diseases Medical sciences Polymerase Chain Reaction - methods RNA isolation RNA, Viral - blood RNA, Viral - genetics RNA, Viral - isolation & purification RNA-Directed DNA Polymerase Sensitivity and Specificity silica RT-PCR Silicon Dioxide Thiocyanates Tropical viral diseases Viral diseases
Online-Zugang:	Volltext
Tags:	Tag hinzufügen Keine Tags, Fügen Sie den ersten Tag hinzu!

Beschreibung
Zusammenfassung:	A rapid, simple and efficient single‐tube procedure is described for the isolation of dengue virus RNA from small amount of serum (10 μl) 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:
ISSN:	0146-6615 1096-9071
DOI:	10.1002/jmv.1890400211