Rapid detection and quantification of Chikungunya virus by a one-step reverse transcription polymerase chain reaction real-time assay.
Chikungunya virus (CHIKV) is a mosquito-transmitted alphavirus. A large outbreak of CHIKV disease occurred in 2005 in the Indian Ocean Islands. Many cases have been imported in European countries. Laboratory confirmation of suspected cases is mandatory for control measures during an outbreak. We report a novel, real-time, reverse transcription-polymerase chain reaction (RT-PCR) for the nonstructural protein 1 region that can quantify a wide range of viral RNA concentrations. This assay was validated by in vitro experiments in which interferon-alpha, a well-known virus inhibitor, showed a dose-dependent inhibition of virus replication on Vero cells that was assessed by viral infectivity and viral RNA production. This new real-time RT-PCR was used to measure viral load in serum samples from cases recently imported to Italy, and may be a useful tool in rapid detection of CHIKV and monitoring the extent of viral replication in patients.
Carletti
Fabrizio
F
National Institute for Infectious Diseases L. Spallanzani, Rome, Italy.
Bordi
Licia
L
Chiappini
Roberta
R
Ippolito
Giuseppe
G
Sciarrone
Maria R
MR
Capobianchi
Maria R
MR
Di Caro
Antonino
A
Castilletti
Concetta
C
D000328:Adult; D018354:Alphavirus Infections / Q000097:blood / Q000821:virology; D001483:Base Sequence; D002646:Chikungunya virus / Q000235:genetics* / Q000302:isolation & purification*; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D012367:RNA, Viral / Q000032:analysis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D019562:Viral Load