Volume 87, Issue 5
  • ISSN: 0002-9637
  • E-ISSN: 1476-1645



Both Chikungunya and Dengue virus belong to the acute arthropod-borne viruses. Because of the lack of specific symptoms, it is difficult to distinguish the two infections based on clinical manifestations. To identify and quantitatively detect Chikungunya and Dengue viruses, a real-time accelerated reverse-transcription-loop-mediated isothermal amplification (RT-LAMP) platform was developed, and 26-confirmed RNA samples, 42 suspects, and 18 healthy serum samples were evaluated by the method. The RT-polymerase chain reaction (PCR) and cDNA sequencing were used as references. The results showed that it could identify the Chikungunya and Dengue virus RNA correctly in all antibody-positive samples within 1 hour, without any cross-reactions. The virus load of the positive samples was quantitatively detected with a turbidimeter. The sensitivity was 100% and specificity was 95.25%. The findings indicate that the RT-LAMP is an effective method for rapid quantity detection of Chikungunya virus and Dengue virus in serum samples with convenient operation, high specificity, and high sensitivity.


Article metrics loading...

Loading full text...

Full text loading...



  1. Ross R, , 1956. The Newala epidemic: III. The virus: isolation, pathogenic properties and relationship to the epidemic. J Hyg (Lond) 54: 177191.[Crossref]
  2. Kelvin AA, , 2011. Outbreak of chikungunya in the Republic of Congo and the global picture. J Infect Developing Ctries 5: 441444.
  3. Schuffenecker I, Iteman I, Michault A, Murri S, Frangeul L, Vaney MC, Lavenir R, Pardigon N, Reynes JM, Pettinelli F, , 2006. Genome microevolution of chikungunya viruses causing the Indian Ocean outbreak. PLoS Med 3: e263.[Crossref]
  4. Ravi V, , 2006. Re-emergence of chikungunya virus in India. Indian J Med Microbiol 24: 8384.[Crossref]
  5. Leo YS, Chow AL, Tan LK, Lye DC, Lin L, Ng LC, , 2009. Chikungunya outbreak, Singapore, 2008. Emerg Infect Dis 15: 836837.[Crossref]
  6. Tsetsarkin KA, Chen R, Leal G, Forrester N, Higgs S, Huang J, Weaver SC, , 2011. Chikungunya virus emergence is constrained in Asia by lineage-specific adaptive landscapes. Proc Natl Acad Sci USA 108: 78727877.[Crossref]
  7. Simon F, Javelle E, Oliver M, Leparc-Goffart I, Marimoutou C, , 2011. Chikungunya virus infection. Curr Infect Dis Rep 13: 218228.[Crossref]
  8. Gubler DJ, , 2002. The global emergence/resurgence of arboviral diseases as public health problems. Arch Med Res 33: 330342.[Crossref]
  9. Saxena P, Dash PK, Santhosh S, Shrivastava A, Parida M, Rao PV, , 2008. Development and evaluation of one step single tube multiplex RT-PCR for rapid detection and typing of dengue viruses. Virol J 5: 20.[Crossref]
  10. Charrel RN, de Lamballerie X, Raoult D, , 2007. Chikungunya outbreaks—the globalization of vector-borne diseases. N Engl J Med 356: 769771.[Crossref]
  11. Lakshmi V, Neeraja M, Subbalaxmi MV, Parida MM, Dash PK, Santhosh SR, Rao PV, , 2008. Clinical features and molecular diagnosis of chikungunya fever from South India. Clin Infect Dis 46: 14361442.[Crossref]
  12. Parola P, de Lamballerie X, Jourdan J, Rovery C, Vaillant V, Minodier P, Brouqui P, Flahault A, Raoult D, Charrel RN, , 2006. Novel chikungunya virus variant in travelers returning from Indian Ocean islands. Emerg Infect Dis 12: 14931498.[Crossref]
  13. Karabatsos N, , 1975. Antigenic relationships of group A arboviruses by plaque reduction neutralization testing. Am J Trop Med Hyg 24: 527532.
  14. Powers AM, Brault AC, Tesh RB, Weaver SC, , 2000. Re-emergence of chikungunya and o'nyong-nyong viruses: evidence for distinct geographical lineages and distant evolutionary relationships. J Gen Virol 81: 471479.[Crossref]
  15. Telles J-N, Le Roux K, Grivard P, Vernet G, Michault A, , 2009. Evaluation of real-time nucleic acid sequence-based amplification for detection of chikungunya virus in clinical samples. J Med Microbiol 58: 11681172.[Crossref]
  16. Hayashi N, Arai R, Tada S, Taguchi H, Ogawa Y, , 2007. Detection and identification of Brettanomyces/Dekkera sp. yeasts with a loop-mediated isothermal amplification method. Food Microbiol 24: 778785.[Crossref]
  17. Ohori A, Endo S, Sano A, Yokoyama K, Yarita K, Yamaguchi M, Kamei K, Miyaji M, Nishimura K, , 2006. Rapid identification of Ochroconis gallopava by a loop-mediated isothermal amplification (LAMP) method. Vet Microbiol 114: 359365.[Crossref]
  18. Notomi T, Okayama H, Masubuchi H, Yonekawa T, Watanabe K, Amino N, Hase T, , 2000. Loop-mediated isothermal amplification of DNA. Nucleic Acids Res 28: e63.[Crossref]
  19. Mori Y, Nagamine K, Tomita N, Notomi T, , 2001. Detection of loop-mediated isothermal amplification reaction by turbidity derived from magnesium pyrophosphate formation. Biochem Biophys Res Commun 289: 150154.[Crossref]
  20. Siyi C, Beilei G, , 2010. Development of a toxR-based loop-mediated isothermal amplification assay for detecting Vibrio parahaemolyticus . BMC Microbiol 10: 41.[Crossref]
  21. Zhao X, Wang L, Chu J, Li Y, Xu Z, Li L, Shirtliff ME, He X, Liu Y, , 2010. Rapid detection of Vibrio parahaemolyticus strains and virulent factors by loop-mediated isothermal amplification assays. Food Sci Biotechnol 19: 11911197.[Crossref]
  22. Bonilauri P, Bellini R, Calzolari M, Angelini R, Venturi L, Fallacara F, Cordioli P, Angelini P, Venturelli C, Merialdi G, , 2008. Chikungunya virus in Aedes albopictus, Italy. Emerg Infect Dis 14: 852.[Crossref]
  23. Bai Z, Liu L, Tu Z, Yao L, Liu J, Xu B, Tang B, Wan Y, Fang M, , 2008. Real-time PCR for detecting circulating dengue virus in the Guangdong Province of China in 2006. J Med Microbiol 57: 15471552.[Crossref]
  24. Zheng K, Li J, Zhang Q, Liang M, Li C, Lin M, Huang J, Li H, Xiang D, Wang N, , 2008. Genetic analysis of chikungunya viruses imported to mainland China in 2008. Virol J 7: 8.[Crossref]
  25. Coker RJ, Hunter BM, Rudge JW, Liverani M, Hanvoravongchai P, , 2011. Emerging infectious diseases in southeast Asia: regional challenges to control. Lancet 377: 599609.[Crossref]
  26. Rezza G, Nicoletti L, Angelini R, Romi R, Finarelli A, Panning M, Cordioli P, Fortuna C, Boros S, Magurano F, , 2007. Infection with chikungunya virus in Italy: an outbreak in a temperate region. Lancet 370: 18401846.[Crossref]
  27. Ravi V, , 2006. Re-emergence of chikungunya virus in India. Indian J Med Microbiol 24: 8384.[Crossref]
  28. Laras K, Sukri NC, Larasati RP, Bangs MJ, Kosim R, , 2005. Tracking the re-emergence of epidemic chikungunya virus in Indonesia. Trans R Soc Trop Med Hyg 99: 128141.[Crossref]
  29. Powers AM, Brault AC, Tesh RB, Weaver SC, , 2000. Re-emergence of chikungunya and o'nyong-nyong viruses: evidence for distinct geographical lineages and distant evolutionary relationships. J Gen Virol 81: 471479.[Crossref]
  30. Parida MM, Santhosh SR, Dash PK, Tripathi NK, Lakshmi V, Mamidi N, Shrivastva A, Gupta N, Saxena P, Babu JP, Rao PV, Morita K, , 2007. Rapid and real-time detection of chikungunya virus by reverse transcription loop-mediated isothermal amplification assay. J Clin Microbiol 45: 351357.[Crossref]
  31. Xu G, Dong H, Shi N, Liu S, Zhou A, Cheng Z, Chen G, Liu J, Fang T, Zhang H, , 2007. An outbreak of dengue virus serotype 1 infection in Cixi, Ningbo, People's Republic of China, 2004, associated with a traveler from Thailand and high density of Aedes albopictus . Am J Trop Med Hyg 76: 11821188.
  32. Lu L, Lin H, Tian L, Yang W, Sun J, Liu Q, , 2009. Time series analysis of dengue fever and weather in Guangzhou, China. BMC Public Health 9: 395.[Crossref]
  33. Fan W, Yu S, Cosgriff TM, , 1989. The reemergence of dengue in China. Rev Infect Dis 11: S847S853.[Crossref]
  34. Qiu FX, Chen QQ, Ho QY, Chen WZ, Zhao ZG, Zhao BW, , 1991. The first epidemic of dengue hemorrhagic fever in the People's Republic of China. Am J Trop Med Hyg 44: 364370.
  35. Qiu FX, Gubler DJ, Liu JC, Chen QQ, , 1993. Dengue in China: a clinical review. Bull World Health Organ 71: 349359.
  36. Gould E, Gallian P, De Lamballerie X, Charrel R, , 2010. First cases of autochthonous dengue fever and chikungunya fever in France: from bad dream to reality! Clin Microbiol Infect 16: 17021704.[Crossref]
  37. Iwamoto T, Sonobe T, Hayashi K, , 2003. Loop-mediated isothermal amplification for direct detection of Mycobacterium tuberculosis complex, M. avium, and M. intracellulare in sputum samples. J Clin Microbiol 41: 26162622.[Crossref]
  38. Bonizzoni M, Afrane Y, Yan G, , 2009. Loop-mediated isothermal amplification (LAMP) for rapid identification of Anopheles gambiae and Anopheles arabiensis mosquitoes. Am J Trop Med Hyg 81: 10301034.[Crossref]
  39. Inacio J, Flores O, Spencer-Martins I, , 2008. Efficient identification of clinically relevant Candida yeast species by use of an assay combining panfungal loop-mediated isothermal DNA amplification with hybridization to species-specific oligonucleotide probes. J Clin Microbiol 46: 713720.[Crossref]
  40. Parida M, Horioke K, Ishida H, Dash PK, Saxena P, Jana AM, Islam MA, 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: 28952903.[Crossref]

Data & Media loading...

  • Received : 18 Nov 2011
  • Accepted : 01 Aug 2012

Most Cited This Month

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error