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Journal of Clinical Microbiology, June 2007, p. 1893-1897, Vol. 45, No. 6
0095-1137/07/$08.00+0     doi:10.1128/JCM.00065-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Evaluation of a Commercial Real-Time PCR Kit for Detection of Dengue Virus in Samples Collected during an Outbreak in Goiânia, Central Brazil, in 2005

José Eduardo Levi,^1* Adriana Fumie Tateno,¹ Adriana Freire Machado,¹ Débora Camillo Ramalho,¹ Vanda Akico Ueda Fick de Souza,¹ Adriana Oliveira Guilarde,² Valéria Christina de Rezende Feres,³ Celina Maria Turchi Martelli,² Marília Dalva Turchi,² João Bosco Siqueira Jr.,² and Cláudio Sérgio Pannuti¹

Laboratório de Virologia, LIM-HC da FMUSP e Instituto de Medicina Tropical de São Paulo, Universidade de São Paulo, São Paulo,¹ Instituto de Patologia Tropical e Saúde Pública (IPTSP), Universidade Federal de Goiás,² LACEN-Go, Secretaria de Estado da Saúde de Goiás, Goiânia, Brazil³

Received 9 January 2007/ Returned for modification 19 February 2007/ Accepted 25 March 2007

In the past 2 decades, dengue has reemerged in Brazil as a significant public health problem. Clinicians demand a diagnostic test with high sensitivity that is applicable during the early symptomatic phase. We aimed to test two distinct molecular methods on samples from suspected dengue cases during an outbreak in Central Brazil. Acute-phase serum specimens from 254 patients suspected of having dengue were collected during 2005 in the city of Goiânia, Central Brazil. Samples were blindly evaluated by real-time and multiplex PCR in addition to routine immunoglobulin M serology and virus culture. Overall, acute dengue was confirmed by serology, multiplex PCR, or virus isolation for 80% of patients (203/254). Another four patients presented real-time PCR-positive results as the unique marker of dengue. Higher real-time PCR positivity levels and viral loads were observed in the early symptomatic phase of disease (5 days) than after this period. Multiplex and real-time PCR assays presented a high kappa agreement (0.85). According to multiplex PCR, 60 samples harbored dengue virus type 3 (DEN-3), 4 samples harbored DEN-2, and 1 sample displayed a pattern compatible with a double infection with DEN-2 and -3. The dengue virus real-time kit was found to be practical and adjustable for high throughput, to display the best performance in the early symptomatic phase of dengue cases, and to be valuable for confirming dengue diagnosis in a timely manner.

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* Corresponding author. Mailing address: Virology Lab, Institute of Tropical Medicine, University of São Paulo, Rua Dr. Enéas de Carvalho Aguiar 470, 2^o andar, CEP 05403-000, São Paulo-SP, Brazil. Phone: 55-11-3062 2645. Fax: 55-11-3063 2659. E-mail: dudilevi{at}usp.br

Published ahead of print on 4 April 2007.

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Journal of Clinical Microbiology, June 2007, p. 1893-1897, Vol. 45, No. 6
0095-1137/07/$08.00+0     doi:10.1128/JCM.00065-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

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