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 Wang, W.-K.
Right arrow Articles by King, C.-C.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Wang, W.-K.
Right arrow Articles by King, C.-C.

 Previous Article  |  Next Article 

Journal of Clinical Microbiology, December 2002, p. 4472-4478, Vol. 40, No. 12
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.12.4472-4478.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

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

Wei-Kung Wang,1* Tzu-Ling Sung,1 Yu-Chen Tsai,1 Chuan-Liang Kao,2 Shu-Mei Chang,3 and Chwan-Chuen King4

Institute of Microbiology,1 Graduate Institute of Medical Technology,2 College of Medicine, and Institute of Epidemiology, College of Public Health,4 National Taiwan University, Taipei, and The Yuan General Hospital, Kaohsiung, Taiwan3

Received 24 June 2002/ Returned for modification 8 August 2002/ Accepted 20 September 2002

While dengue virus is thought to replicate in mononuclear phagocytic cells in vivo, attempts to detect it in peripheral blood mononuclear cells (PBMC) by virus isolation or antigen detection have had variable and generally low rates. In this study, we developed a reverse transcription (RT)-real-time PCR assay to quantify positive- and negative-sense RNA of dengue virus type 2 within the cells. The assay includes an RT step using either sense or antisense primer followed by a real-time PCR step using the designed primers and probe, which target a capsid region highly conserved in dengue virus type 2 strains. It can be used to monitor the dynamic change of intracellular dengue virus RNA species during the course of infection. When this assay is employed in quantification of dengue virus RNA species in PBMC from 10 patients infected with dengue virus type 2, both positive- and negative-sense dengue RNA can be detected, indicating that dengue virus is actively replicating in PBMC in vivo. Moreover, the amounts of negative-sense dengue virus RNA in PBMC correlate very well with the viral load of dengue virus in plasma, suggesting that quantification of negative-sense dengue virus RNA in PBMC may provide another indicator of dengue virus replication in vivo. Use of this convenient, sensitive, and accurate method of quantification in clinical samples from patients with different disease severity would further our understanding of the pathogenesis of dengue.

* Corresponding author. Mailing address: Institute of Microbiology, College of Medicine, National Taiwan University, No. 1 Sec. 1 Jen-Ai Rd., Taipei, Taiwan. Phone: 886-2-2312-3456, ext. 8286. Fax: 886-2-2391-5293. E-mail: wwang60{at}yahoo.com.

Journal of Clinical Microbiology, December 2002, p. 4472-4478, Vol. 40, No. 12
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.12.4472-4478.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Anwar, A., Wan, G., Chua, K.-B., August, J. T., Too, H.-P. (2009). Evaluation of Pre-Analytical Variables in the Quantification of Dengue Virus by Real-Time Polymerase Chain Reaction. J. Mol. Diagn. 11: 537-542 [Abstract] [Full Text]  
  • Kroschewski, H., Sagripanti, J.-L., Davidson, A. D. (2009). Identification of amino acids in the dengue virus type 2 envelope glycoprotein critical to virus infectivity. J. Gen. Virol. 90: 2457-2461 [Abstract] [Full Text]  
  • Mota, J., Rico-Hesse, R. (2009). Humanized Mice Show Clinical Signs of Dengue Fever according to Infecting Virus Genotype. J. Virol. 83: 8638-8645 [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]  
  • Chen, H.-C., Hofman, F. M., Kung, J. T., Lin, Y.-D., Wu-Hsieh, B. A. (2007). Both Virus and Tumor Necrosis Factor Alpha Are Critical for Endothelium Damage in a Mouse Model of Dengue Virus-Induced Hemorrhage. J. Virol. 81: 5518-5526 [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]  
  • ANDERSON, J. R., RICO-HESSE, R. (2006). AEDES AEGYPTI VECTORIAL CAPACITY IS DETERMINED BY THE INFECTING GENOTYPE OF DENGUE VIRUS. Am J Trop Med Hyg 75: 886-892 [Abstract] [Full Text]  
  • Kang, X., Yang, B.-a., Hu, Y., Zhao, H., Xiong, W., Yang, Y., Si, B., Zhu, Q. (2006). Human Neutralizing Fab Molecules against Severe Acute Respiratory Syndrome Coronavirus Generated by Phage Display.. CVI 13: 953-957 [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]  
  • RICHARDSON, J., MOLINA-CRUZ, A., SALAZAR, M. I., BLACK, W. IV (2006). QUANTITATIVE ANALYSIS OF DENGUE-2 VIRUS RNA DURING THE EXTRINSIC INCUBATION PERIOD IN INDIVIDUAL AEDES AEGYPTI. Am J Trop Med Hyg 74: 132-141 [Abstract] [Full Text]  
  • Bente, D. A., Melkus, M. W., Garcia, J. V., Rico-Hesse, R. (2005). Dengue Fever in Humanized NOD/SCID Mice. J. Virol. 79: 13797-13799 [Abstract] [Full Text]  
  • Jones, M., Davidson, A., Hibbert, L., Gruenwald, P., Schlaak, J., Ball, S., Foster, G. R., Jacobs, M. (2005). Dengue Virus Inhibits Alpha Interferon Signaling by Reducing STAT2 Expression. J. Virol. 79: 5414-5420 [Abstract] [Full Text]  
  • MOLINA-CRUZ, A., GUPTA, L., RICHARDSON, J., BENNETT, K., BLACK, W. IV, BARILLAS-MURY, C. (2005). EFFECT OF MOSQUITO MIDGUT TRYPSIN ACTIVITY ON DENGUE-2 VIRUS INFECTION AND DISSEMINATION IN AEDES AEGYPTI. Am J Trop Med Hyg 72: 631-637 [Abstract] [Full Text]  
  • Shu, P.-Y., Huang, J.-H. (2004). Current Advances in Dengue Diagnosis. CVI 11: 642-650 [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»