Volume 101, Issue 2
  • ISSN: 0002-9637
  • E-ISSN: 1476-1645



Molecular epidemiological data are key for dengue outbreak characterization and preparedness. However, sparse (DENV) molecular information is available in Laos because of limited resources. In this proof-of-concept study, we evaluated whether DENV1 RNA extracted from rapid diagnostic tests (RDTs) could be amplified and sequenced. The protocol for envelope gene amplification from RNA purified from RDTs was first assessed using viral isolate dilutions then conducted using 14 dengue patient sera. Envelope gene amplification was successful from patient sera with high virus titer, as was sequencing but with lower efficiency. Hence, based on our results, RDTs can be a source of DENV1 RNA for subsequent envelope gene amplification and sequencing. This is a promising tool for collecting molecular epidemiology data from rural dengue-endemic areas. However, further investigations are needed to improve assay efficiency and to assess this tool’s level of efficacy on a larger scale in the field.

[open-access] This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.


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  1. World Health Organization (WHO), 2017. Dengue and severe dengue . Available at http://www.who.int/mediacentre/factsheets/fs117/en/. Accessed July 20, 2017.
  2. Bhatt S, 2013. The global distribution and burden of dengue. Nature 496: 504507. [Google Scholar]
  3. Khampapongpane B, Lewis HC, Ketmayoon P, Phonekeo D, Somoulay V, Khamsing A, Phengxay M, Sisouk T, Vongphrachanh P, Bryant JE, , 2014. National dengue surveillance in the Lao People’s Democratic Republic, 2006-2012: epidemiological and laboratory findings. West Pac Surveill Response J 5: 713. [Google Scholar]
  4. Dubot-Pérès A, 2013. An epidemic of dengue-1 in a remote village in rural Laos. PLoS Negl Trop Dis 7: e2360. [Google Scholar]
  5. Castonguay-Vanier J, 2018. Molecular epidemiology of dengue viruses in three provinces of Lao PDR, 2006–2010. PLoS Negl Trop Dis 12: e0006203. [Google Scholar]
  6. Mayxay M, 2011. Predictive diagnostic value of the tourniquet test for the diagnosis of dengue infection in adults: accuracy of the tourniquet test for the diagnosis of dengue. Trop Med Int Health 16: 127133. [Google Scholar]
  7. Nisalak A, Endy TP, Nimmannitya S, Kalayanarooj S, Thisayakorn U, Scott RM, Burke DS, Hoke CH, Innis BL, Vaughn DW, , 2003. Serotype-specific dengue virus circulation and dengue disease in Bangkok, Thailand from 1973 to 1999. Am J Trop Med Hyg 68: 191202. [Google Scholar]
  8. Balmaseda A, 2006. Serotype-specific differences in clinical manifestations of dengue. Am J Trop Med Hyg 74: 449456. [Google Scholar]
  9. Dash PK, Parida MM, Saxena P, Abhyankar A, Singh CP, Tewari KN, Jana AM, Sekhar K, Rao PVL, , 2006. Reemergence of dengue virus type-3 (subtype-III) in India: implications for increased incidence of DHF & DSS. Virol J 3: 55. [Google Scholar]
  10. Lee K-S, Lo S, Tan SS-Y, Chua R, Tan L-K, Xu H, Ng L-C, , 2012. Dengue virus surveillance in Singapore reveals high viral diversity through multiple introductions and in situ evolution. Infect Genet Evol 12: 7785. [Google Scholar]
  11. Lao M, 2014. Co-circulation of dengue virus type 3 genotypes in Vientiane capital, Lao P.D.R. PLoS One 9: e115569. [Google Scholar]
  12. Fukunaga T, Phommasack B, Bounlu K, Saito M, Tadano M, Makino Y, Kanemura K, Arakaki S, Shinjo M, , 1993. Epidemiological situation of dengue infection in Lao P.D.R. Trop Med 35: 219227. [Google Scholar]
  13. Peyerl-Hoffmann G, Schwöbel B, Jordan S, Vamisaveth V, Phetsouvanh R, Christophel EM, Phompida S, Sonnenburg FV, Jelinek T, , 2004. Serological investigation of the prevalence of anti-dengue IgM and IgG antibodies in Attapeu Province, South Laos. Clin Microbiol Infect 10: 181184. [Google Scholar]
  14. Yang H, 2015. Molecular characterization and viral origin of the first dengue outbreak in Xishuangbanna, Yunnan Province, China, 2013. Am J Trop Med Hyg 93: 390393. [Google Scholar]
  15. Vongsouvath M, 2016. Using rapid diagnostic tests as a source of viral RNA for dengue serotyping by RT-PCR - a novel epidemiological tool. PLoS Negl Trop Dis 10: e0004704. [Google Scholar]
  16. Sengvilaipaseuth O, 2017. Temperature of a dengue rapid diagnostic test under tropical climatic conditions: a follow up study. PLoS One 12: e0170359. [Google Scholar]
  17. Phommasone K, Sengvilaipaseuth O, de Lamballerie X, Vongsouvath M, Phonemixay O, Blacksell SD, Newton PN, Dubot-Pérès A, , 2015. Temperature and the field stability of a dengue rapid diagnostic test in the tropics. Am J Trop Med Hyg 93: 3339. [Google Scholar]
  18. Tang Y, Quintana M, Zhang C, Li T, Sauls D, Putnak R, Alvarado SC, Castro DJ, Matute ML, Lynch JA, , 2011. Letter to the editor: molecular genotyping of Dengue Virus Types 2 and 4 from the Guatemalan and Honduran Epidemics of 2007 using the envelope glycoprotein gene. Virus Genes 42: 200203. [Google Scholar]
  19. Leparc-Goffart I, Baragatti M, Temmam S, Tuiskunen A, Moureau G, Charrel R, de Lamballerie X, , 2009. Development and validation of real-time one-step reverse transcription-PCR for the detection and typing of dengue viruses. J Clin Virol 45: 6166. [Google Scholar]
  20. Kumar S, Stecher G, Tamura K, , 2016. MEGA7: molecular evolutionary genetics analysis version 7.0 for bigger datasets. Mol Biol Evol 33: 18701874. [Google Scholar]

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  • Received : 17 Oct 2018
  • Accepted : 06 May 2019
  • Published online : 24 Jun 2019

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