Development of a TaqMan RT-PCR assay without RNA extraction step for the detection and quantification of African Chikungunya viruses.
Chikungunya virus (CHIKV), a member of the alphavirus genus, is of considerable public health concern in Southeast Asian and African countries. However, despite serological evidence, the diagnosis of this arthropod-borne human disease is confirmed infrequently and 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, and 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 and cell culture supernatants. In this study, a specific and 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.
Pastorino
Boris
B
Unité de virologie tropicale, Laboratoire associé au Centre national de référence pour les arbovirus, Institut de médecine tropicale du service de santé des armées, BP 46, Parc du Pharo, 13998 Marseille Armées, France. publi.viro@laposte.net
Bessaud
Maël
M
Grandadam
Marc
M
Murri
Severine
S
Tolou
Hugues J
HJ
Peyrefitte
Christophe N
CN
D001483:Base Sequence; D002646:Chikungunya virus / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D012367:RNA, Viral / Q000302:isolation & purification*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
D012367:RNA, Viral