1921
Volume 79, Issue 4
  • ISSN: 0002-9637
  • E-ISSN: 1476-1645

Abstract

We have systematically evaluated a dry-format, quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) assay developed by Tetracore Inc. for the Cepheid SmartCycler platform to facilitate rapid diagnosis of dengue virus infections. A panel of related flaviviruses was used to evaluate the clinical specificity of the assay, and it was found to be specific to dengue. Eighty-one clinical samples previously confirmed dengue positive by virus isolation, along with 25 dengue negative control specimens were used to validate this new diagnostic assay. Using these clinical samples, the assay exhibited 98.77% sensitivity and 100% specificity. Over 85% of the clinical specimen exhibited viral loads ranging from 10 to 10 plaque-forming units per milliliter (PFU/mL). In addition, this dry-format assay is stable at ambient temperatures and requires minimal technical expertise to perform in a small thermocycler platform. These characteristics make it a promising candidate for diagnosis of dengue in mobile laboratories in the field.

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2008-10-01
2017-11-20
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References

  1. Rico-Hesse R, 2003. Microevolution and virulence of dengue viruses. Adv Virus Res 59 : 315–341.
  2. Trent DW, Manske CL, Fox GE, Chu MC, Kliks SC, Monath TP, 1989. The molecular epidemiology of dengue viruses, genetic variation and microevolution. Kurstk E, ed. Applied Virology Research, Virus Variation and Epidemiology. Plenum, NY: Plenum Press.
  3. WHO, Strengthening Implementation of the Global Strategy for Dengue Fever and Dengue Haemorrhagic Fever, Prevention and Control. Report of the informal consultation. Geneva, Switzerland: World Health Organization, 1999.
  4. Gubler DJ, 1998. Dengue and dengue hemorrhagic fever. Clin Microbiol Rev 11 : 480–496.
  5. George R, Lum L, 1997. Clinical spectrum of dengue infection. Gubler DJ, Kuno G, eds. Dengue and Dengue Hemorrhagic Fever. Wallingford, United Kingdom: CAB International, 89–113.
  6. Henchal EA, McCown JM, Seguin MC, Gentry MK, Brandt WE, 1983. Rapid identification of dengue virus isolates by using monoclonal antibodies in an indirect immunofluorescence assay. Am J Trop Med Hyg 32 : 164–169.
  7. Rigau-Perez JD, Gubler DJ, 1997. Dengue: a literature review and case study of travelers from the United States, 1986–1994. J Travel Med 4 : 65–71.
  8. Halstead SB, 1988. Pathogenesis of dengue: challenges to molecular biology. Science 239 : 476–481.
  9. Rigau-Perez JG, Clark GG, Gubler DJ, Reiter P, Sanders EJ, Vorndam AV, 1998. Dengue and dengue haemorrhagic fever. Lancet 352 : 971–977.
  10. Teles FR, Prazeres DM, Lima-Filho JL, 2005. Trends in dengue diagnosis. Rev Med Virol 15 : 287–302.
  11. Shu PY, Huang JH, 2004. Current advances in dengue diagnosis. Clin Diagn Lab Immunol 11 : 642–650.
  12. Kao CL, King CC, Chao DY, Wu HL, Chang GJ, 2005. Laboratory diagnosis of dengue virus infection: current and future perspectives in clinical diagnosis and public health. J Microbiol Immunol Infect 38 : 5–16.
  13. Callahan JD, Wu SJ, Dion-Schultz A, Mangold BE, Peruski LF, Watts DM, Porter KR, Murphy GR, Suharyono W, King CC, Hayes CG, Temenak JJ, 2001. Development and evaluation of serotype- and group-specific fluorogenic reverse transcriptase PCR (TaqMan) assays for dengue virus. J Clin Microbiol 39 : 4119–4124.
  14. Houng HS, Chung-Ming Chen R, Vaughn DW, Kanesathasan N, 2001. Development of a fluorogenic RT-PCR system for quantitative identification of dengue virus serotypes 1–4 using conserved and serotype-specific 3′ noncoding sequences. J Virol Methods 95 : 19–32.
  15. Houng HS, Hritz D, Kanesathasan N, 2000. Quantitative detection of dengue 2 virus using fluorogenic RT-PCR based on 3′-noncoding sequence. J Virol Methods 86 : 1–11.
  16. Lanciotti RS, Calisher CH, Gubler DJ, Chang GJ, Vorndam AV, 1992. Rapid detection and typing of dengue viruses from clinical samples by using reverse transcriptase-polymerase chain reaction. J Clin Microbiol 30 : 545–551.
  17. Lanciotti RS, 2003. Molecular amplification assays for the detection of flaviviruses. Adv Virus Res 61 : 67–99.
  18. Henchal EA, McCown JM, Seguin MC, Gentry MK, Brandt WE, 1983. Rapid identification of dengue virus isolates by using monoclonal antibodies in an indirect immunofluorescence assay. Am J Trop Med Hyg 32 : 164–169.
  19. Cheng ZJ, Hu LH, Fu WR, Li YR, 2007. Rapid quantification of hepatitis B virus DNA by direct real-time PCR from serum without DNA extraction. J Med Microbiol 56 : 766–771.
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  • Received : 20 Nov 2007
  • Accepted : 18 Jun 2008

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