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Advances in Virology
Volume 2011, Article ID 514681, 9 pages
http://dx.doi.org/10.1155/2011/514681
Research Article

Internally Controlled, Generic Real-Time PCR for Quantification and Multiplex Real-Time PCR with Serotype-Specific Probes for Serotyping of Dengue Virus Infections

1Royal Tropical Institute, KIT Biomedical Research, Meibergdreef 39, 1105 AZ Amsterdam, The Netherlands
2Division of Infectious Diseases, Academic Medical Center, Tropical Medicine & AIDS, Meibergdreef 9, 1105 AZ Amsterdam, The Netherlands
3Center for Infection and Immunity (CINIMA), Academic Medical Center, University of Amsterdam, 1105 AZ Amsterdam, The Netherlands
4Division of Microbiology, Cho Ray Hospital, 201B Nguyen Chi Thanh Street, District 5, Ho Chi Minh City, Vietnam
5Division of Medical Microbiology, Academic Medical Center, Section of Clinical Virology, Meibergdreef 9, 1105 AZ Amsterdam, The Netherlands

Received 25 July 2011; Accepted 10 September 2011

Academic Editor: Jay C. Brown

Copyright © 2011 Sandra Menting et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Dengue has become a global public health problem and a sensitive diagnostic test for early phase detection can be life saving. An internally controlled, generic real-time PCR was developed and validated by testing serial dilutions of a DENV positive control RNA in the presence of a fixed amount of IC with results showing a good linearity ( ) and a LOD of at least  copies/mL. Application of the generic PCR on 136 patient samples revealed a sensitivity of 95.8% and specificity of 100%. A newly developed multiplex real-time PCR with serotype-specific probes allowed the serotyping of DENV for 80 out of 92 (87%) generic real-time PCR positive patients. Combined these real-time PCRs offer a convenient diagnostic tool for the sensitive and specific quantification of DENV in clinical specimens with the possibility for serotyping.