Development of a TaqMan RT-PCR assay without RNA extraction step for the detection and quantification of African Chikungunya viruses
Autor(es): Pastorino Boris, Bessaud Maël, Grandadam Marc, Murri Severine, Tolou Hugues J, Peyrefitte Christophe N
Resumo: Chikungunya virus (CHIKV), a member of the alphavirus genus, is of considerable public health concern in Southeast Asian - African countries. However, despite serological evidence, the diagnosis of this arthropod-borne human disease is confirmed infrequently - needs to be improved. In fact, illness caused by CHIKV can be confused with diseases such as dengue or yellow fever, based on the similarity of the symptoms, - laboratory confirmation of suspected cases is required to launch control measures during an epidemic. Moreover, no quantitative molecular tool is described to study CHIKV replication or detection in clinical samples - cell culture supernatants. In this study, a specific - sensitive CHIKV one-step TaqMan RT-PCR assay was developed as a tool for the diagnosis of African CHIKV as well as a rapid indicator of active infection by quantifying viral load. This study also showed that a simple heat viral RNA release during the reverse transcription step constituted an alternative to the conventional RNA extraction method.
Palavras-Chave: Alphavirus; Chikungunya virus; Heat-release; Direct one-step TaqMan®; RT-PCR
Imprenta: Journal of Virological Methods, v. 124, n. 1-2, p. 65-71, 2005
Identificador do objeto digital: 10.1016/j.jviromet.2004.11.002
Descritores: Chikungunya virus - Arbovirus ; Chikungunya virus - Cell ; Chikungunya virus - Molecular structure ; Chikungunya virus - RNA ; Chikungunya virus - Viral infections ; Chikungunya virus - Molecular methods ; Chikungunya virus - PCR detection ; Chikungunya virus - RT-PCR ; Chikungunya virus - Serological diagnosis ; Chikungunya Virus - Virus ; Chikungunya virus - Molecular screening ; Chikungunya virus - Chikungunya fever ; Chikungunya virus - Dengue ; Chikungunya virus - Epidemic ; Chikungunya virus - Public health
Data de publicação: 2005
