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Am. J. Trop. Med. Hyg., 77(3), 2007, pp. 521-524
Copyright © 2007 by The American Society of Tropical Medicine and Hygiene

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SHORT REPORT


Rapid Detection and Quantification of Chikungunya Virus by a One-Step Reverse Transcription–Polymerase Chain Reaction Real-Time Assay

Fabrizio Carletti, Licia Bordi, Roberta Chiappini, Giuseppe Ippolito, Maria R. Sciarrone, Maria R. Capobianchi, Antonino Di Caro*, AND Concetta Castilletti
National Institute for Infectious Diseases L. Spallanzani, Rome, Italy

 

ABSTRACT

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.

Received March 9, 2007. Accepted for publication May 29, 2007.

Acknowledgments: We thank Francesco Fusco, Vincenzo Puro, and Emanuele Nicastri for providing epidemiologic assistance in tracing patient medical histories.

Financial support: The study was partially supported by grants from the Italian Ministry of Health, Fondi Ricerca Corrente, and Piano di Sostegno Diagnostico Assistenziale ed Epidemiologico alle Emergenze Biologiche sul Territorio Italiano.

* Address correspondence to Antonino Di Caro, Laboratory of Virology, Padiglione Baglivi, National Institute for Infectious Diseases L. Spallanzani, Via Portuense 292, 00149 Rome, Italy. E-mail: dicaro{at}inmi.it

Authors’ addresses: Fabrizio Carletti, Licia Bordi, Roberta Chiappini, Maria R. Sciarrone, Maria R. Capobianchi, Antonino Di Caro, and Concetta Castilletti, Laboratory of Virology, National Institute for Infectious Diseases L. Spallanzani, Rome, Italy, E-mails: carletti{at}inmi.it, lbordi{at}inmi.it, chiappini{at}inmi.it, sciarrone{at}inmi.it, capobianchi{at}inmi.it, dicaro{at}inmi.it, and castilletti{at}inmi.it. Giuseppe Ippolito, Department of Epidemiology, National Institute for Infectious Diseases L. Spallanzani, Rome, Italy, E-mail: ippolito{at}inmi.it.

Reprint requests: Antonino Di Caro Laboratory of Virology, Padiglione Baglivi National Institute for Infectious Diseases L. Spallanzani, Via Portuense 292, 00149 Rome, Italy.






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