This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Harris, E.
Right arrow Articles by Balmaseda, A.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Harris, E.
Right arrow Articles by Balmaseda, A.

 Previous Article  |  Next Article 

Journal of Clinical Microbiology, September 1998, p. 2634-2639, Vol. 36, No. 9
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Typing of Dengue Viruses in Clinical Specimens and Mosquitoes by Single-Tube Multiplex Reverse Transcriptase PCR

Eva Harris,1,* T. Guy Roberts,1 Leila Smith,1 John Selle,1 Laura D. Kramer,2 Sonia Valle,3 Erick Sandoval,3 and Angel Balmaseda3

Program in Molecular Pathogenesis, University of California, San Francisco, San Francisco, California 94143-04221; Center for Vector-Borne Disease Research, University of California, Davis, Davis, California 956162; and Centro Nacional de Diagnóstico y Referencia, Ministerio de Salud, Managua, Nicaragua3

Received 17 February 1998/Returned for modification 15 May 1998/Accepted 8 June 1998

In recent years, dengue viruses (serotypes 1 to 4) have spread throughout tropical regions worldwide. In many places, multiple dengue virus serotypes are circulating concurrently, which may increase the risk for the more severe form of the disease, dengue hemorrhagic fever. For the control and prevention of dengue fever, it is important to rapidly detect and type the virus in clinical samples and mosquitoes. Assays based on reverse transcriptase (RT) PCR (RT-PCR) amplification of dengue viral RNA can offer a rapid, sensitive, and specific approach to the typing of dengue viruses. We have reduced a two-step nested RT-PCR protocol to a single-tube reaction with sensitivity equivalent to that of the two-step protocol (1 to 50 PFU) in order to maximize simplicity and minimize the risk of sample cross-contamination. This assay was also optimized for use with a thermostable RT-polymerase. We designed a plasmid-based internal control that produces a uniquely sized product and can be used to control for both reverse transcription or amplification steps without the risk of generating false-positive results. This single-tube RT-PCR procedure was used to type dengue viruses during the 1995 and 1997-1998 outbreaks in Nicaragua. In addition, an extraction procedure that permits the sensitive detection of viral RNA in pools of up to 50 mosquitoes without PCR inhibition or RNA degradation was developed. This assay should serve as a practical tool for use in countries where dengue fever is endemic, in conjunction with classical methods for surveillance and epidemiology of dengue viruses.

* Corresponding author. Present address: Division of Public Health Biology and Epidemiology, School of Public Health, University of California, Berkeley, 140 Warren Hall, Berkeley, CA 94720. Phone: (510) 643-9773. Fax: (510) 642-6350. E-mail: eharris{at}uclink4.berkeley.edu.

Journal of Clinical Microbiology, September 1998, p. 2634-2639, Vol. 36, No. 9
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Das, S., Pingle, M. R., Munoz-Jordan, J., Rundell, M. S., Rondini, S., Granger, K., Chang, G.-J. J., Kelly, E., Spier, E. G., Larone, D., Spitzer, E., Barany, F., Golightly, L. M. (2008). Detection and Serotyping of Dengue Virus in Serum Samples by Multiplex Reverse Transcriptase PCR-Ligase Detection Reaction Assay. J. Clin. Microbiol. 46: 3276-3284 [Abstract] [Full Text]  
  • Bessoff, K., Delorey, M., Sun, W., Hunsperger, E. (2008). Comparison of Two Commercially Available Dengue Virus (DENV) NS1 Capture Enzyme-Linked Immunosorbent Assays Using a Single Clinical Sample for Diagnosis of Acute DENV Infection. CVI 15: 1513-1518 [Abstract] [Full Text]  
  • Jackson, G. W., McNichols, R. J., Fox, G. E., Willson, R. C. (2008). Toward Universal Flavivirus Identification by Mass Cataloging. J. Mol. Diagn. 10: 135-141 [Abstract] [Full Text]  
  • Levi, J. E., Tateno, A. F., Machado, A. F., Ramalho, D. C., de Souza, V. A. U. F., Guilarde, A. O., de Rezende Feres, V. C., Martelli, C. M. T., Turchi, M. D., Siqueira, J. B. Jr., Pannuti, C. S. (2007). Evaluation of a Commercial Real-Time PCR Kit for Detection of Dengue Virus in Samples Collected during an Outbreak in Goiania, Central Brazil, in 2005. J. Clin. Microbiol. 45: 1893-1897 [Abstract] [Full Text]  
  • Tavakoli, N. P., Tobin, E. H., Wong, S. J., Dupuis, A. P. II, Glasheen, B., Kramer, L. D., Bernard, K. A. (2007). Identification of Dengue Virus in Respiratory Specimens from a Patient Who Had Recently Traveled from a Region Where Dengue Virus Infection Is Endemic. J. Clin. Microbiol. 45: 1523-1527 [Abstract] [Full Text]  
  • Dussart, P., Labeau, B., Lagathu, G., Louis, P., Nunes, M. R. T., Rodrigues, S. G., Storck-Herrmann, C., Cesaire, R., Morvan, J., Flamand, M., Baril, L. (2006). Evaluation of an Enzyme Immunoassay for Detection of Dengue Virus NS1 Antigen in Human Serum. CVI 13: 1185-1189 [Abstract] [Full Text]  
  • Chien, L.-J., Liao, T.-L., Shu, P.-Y., Huang, J.-H., Gubler, D. J., Chang, G.-J. J. (2006). Development of Real-Time Reverse Transcriptase PCR Assays To Detect and Serotype Dengue Viruses. J. Clin. Microbiol. 44: 1295-1304 [Abstract] [Full Text]  
  • MENDEZ, F., BARRETO, M., ARIAS, J. F., RENGIFO, G., MUNOZ, J., BURBANO, M. E., PARRA, B. (2006). HUMAN AND MOSQUITO INFECTIONS BY DENGUE VIRUSES DURING AND AFTER EPIDEMICS IN A DENGUE-ENDEMIC REGION OF COLOMBIA. Am J Trop Med Hyg 74: 678-683 [Abstract] [Full Text]  
  • BALMASEDA, A., HAMMOND, S. N., PEREZ, L., TELLEZ, Y., SABORIO, S. I., MERCADO, J. C., CUADRA, R., ROCHA, J., PEREZ, M. A., SILVA, S., ROCHA, C., HARRIS, E. (2006). SEROTYPE-SPECIFIC DIFFERENCES IN CLINICAL MANIFESTATIONS OF DENGUE.. Am J Trop Med Hyg 74: 449-456 [Abstract] [Full Text]  
  • HAMMOND, S. N., BALMASEDA, A., PEREZ, L., TELLEZ, Y., SABORIO, S. I., MERCADO, J. C., VIDEA, E., RODRIGUEZ, Y., PEREZ, M. A., CUADRA, R., SOLANO, S., ROCHA, J., IDIAQUEZ, W., GONZALEZ, A., HARRIS, E. (2005). DIFFERENCES IN DENGUE SEVERITY IN INFANTS, CHILDREN, AND ADULTS IN A 3-YEAR HOSPITAL-BASED STUDY IN NICARAGUA. Am J Trop Med Hyg 73: 1063-1070 [Abstract] [Full Text]  
  • Lindegren, G., Vene, S., Lundkvist, A., Falk, K. I. (2005). Optimized Diagnosis of Acute Dengue Fever in Swedish Travelers by a Combination of Reverse Transcription-PCR and Immunoglobulin M Detection. J. Clin. Microbiol. 43: 2850-2855 [Abstract] [Full Text]  
  • Parida, M., Horioke, K., Ishida, H., Dash, P. K., Saxena, P., Jana, A. M., Islam, M. A., Inoue, S., Hosaka, N., Morita, K. (2005). Rapid Detection and Differentiation of Dengue Virus Serotypes by a Real-Time Reverse Transcription-Loop-Mediated Isothermal Amplification Assay. J. Clin. Microbiol. 43: 2895-2903 [Abstract] [Full Text]  
  • Bronzoni, R. V. d. M., Baleotti, F. G., Ribeiro Nogueira, R. M., Nunes, M., Moraes Figueiredo, L. T. (2005). Duplex Reverse Transcription-PCR Followed by Nested PCR Assays for Detection and Identification of Brazilian Alphaviruses and Flaviviruses. J. Clin. Microbiol. 43: 696-702 [Abstract] [Full Text]  
  • Coloma, J., Harris, E. (2004). Innovative low cost technologies for biomedical research and diagnosis in developing countries. BMJ 329: 1160-1162 [Full Text]  
  • SANTOS, F. B. D., MIAGOSTOVICH, M. P., NOGUEIRA, R. M. R., SCHATZMAYR, H. G., RILEY, L. W., HARRIS, E. (2004). ANALYSIS OF RECOMBINANT DENGUE VIRUS POLYPEPTIDES FOR DENGUE DIAGNOSIS AND EVALUATION OF THE HUMORAL IMMUNE RESPONSE. Am J Trop Med Hyg 71: 144-152 [Abstract] [Full Text]  
  • Shu, P.-Y., Huang, J.-H. (2004). Current Advances in Dengue Diagnosis. CVI 11: 642-650 [Full Text]  
  • Shu, P.-Y., Chang, S.-F., Kuo, Y.-C., Yueh, Y.-Y., Chien, L.-J., Sue, C.-L., Lin, T.-H., Huang, J.-H. (2003). Development of Group- and Serotype-Specific One-Step SYBR Green I-Based Real-Time Reverse Transcription-PCR Assay for Dengue Virus. J. Clin. Microbiol. 41: 2408-2416 [Abstract] [Full Text]  
  • Balmaseda, A., Guzman, M. G., Hammond, S., Robleto, G., Flores, C., Tellez, Y., Videa, E., Saborio, S., Perez, L., Sandoval, E., Rodriguez, Y., Harris, E. (2003). Diagnosis of Dengue Virus Infection by Detection of Specific Immunoglobulin M (IgM) and IgA Antibodies in Serum and Saliva. CVI 10: 317-322 [Abstract] [Full Text]  
  • Wang, W.-K., Sung, T.-L., Tsai, Y.-C., Kao, C.-L., Chang, S.-M., King, C.-C. (2002). Detection of Dengue Virus Replication in Peripheral Blood Mononuclear Cells from Dengue Virus Type 2-Infected Patients by a Reverse Transcription-Real-Time PCR Assay. J. Clin. Microbiol. 40: 4472-4478 [Abstract] [Full Text]  
  • Wang, W.-K., Lin, S.-R., Lee, C.-M., King, C.-C., Chang, S.-C. (2002). Dengue Type 3 Virus in Plasma Is a Population of Closely Related Genomes: Quasispecies. J. Virol. 76: 4662-4665 [Abstract] [Full Text]  
  • Kao, C.-L., Wu, M.-C., Chiu, Y.-H., Lin, J.-L., Wu, Y.-C., Yueh, Y.-Y., Chen, L.-K., Shaio, M.-F., King, C.-C. (2001). Flow Cytometry Compared with Indirect Immunofluorescence for Rapid Detection of Dengue Virus Type 1 after Amplification in Tissue Culture. J. Clin. Microbiol. 39: 3672-3677 [Abstract] [Full Text]  
  • Wu, S.-J. L., Lee, E. M., Putvatana, R., Shurtliff, R. N., Porter, K. R., Suharyono, W., Watts, D. M., King, C.-C., Murphy, G. S., Hayes, C. G., Romano, J. W. (2001). Detection of Dengue Viral RNA Using a Nucleic Acid Sequence-Based Amplification Assay. J. Clin. Microbiol. 39: 2794-2798 [Abstract] [Full Text]  
  • Benjeddou, M., Leat, N., Allsopp, M., Davison, S. (2001). Detection of Acute Bee Paralysis Virus and Black Queen Cell Virus from Honeybees by Reverse Transcriptase PCR. Appl. Environ. Microbiol. 67: 2384-2387 [Abstract] [Full Text]  
  • Wang, W.-K., Lee, C.-N., Kao, C.-L., Lin, Y.-L., King, C.-C. (2000). Quantitative Competitive Reverse Transcription-PCR for Quantification of Dengue Virus RNA. J. Clin. Microbiol. 38: 3306-3310 [Abstract] [Full Text]  
  • Diamond, M. S., Edgil, D., Roberts, T. G., Lu, B., Harris, E. (2000). Infection of Human Cells by Dengue Virus Is Modulated by Different Cell Types and Viral Strains. J. Virol. 74: 7814-7823 [Abstract] [Full Text]  
  • Diamond, M. S., Roberts, T. G., Edgil, D., Lu, B., Ernst, J., Harris, E. (2000). Modulation of Dengue Virus Infection in Human Cells by Alpha, Beta, and Gamma Interferons. J. Virol. 74: 4957-4966 [Abstract] [Full Text]  
  • Miagostovich, M. P., dos Santos, F. B., Gutiérrez, C. M., Riley, L. W., Harris, E. (2000). Rapid Subtyping of Dengue Virus Serotypes 1 and 4 by Restriction Site-Specific PCR. J. Clin. Microbiol. 38: 1286-1289 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»