Skip to main content

Advertisement

SpringerLink
Log in

Development of a multiplex real-time RT-PCR assay for simultaneous detection of dengue and chikungunya viruses

  • Brief Report
  • Published:
Archives of Virology Aims and scope Submit manuscript

Abstract

Dengue and chikungunya viruses co-circulate and cause infections that start with similar symptoms but progress to radically different outcomes. Therefore, an early diagnostic test that can differentiate between the two is needed. A single-step multiplex real-time RT-PCR assay was developed that can simultaneously detect and quantitate RNA of all dengue virus (DENV) serotypes and chikungunya virus (CHIKV). The sensitivity was 100 % for DENV and 95.8 % for CHIKV, whilst the specificity was 100 % for both viruses when compared with conventional RT-PCR. The detection limit ranged from 1 to 50 plaque-forming units. The assay was successfully used for differential diagnosis of dengue and chikungunya in Pune, where the viruses co-circulate.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2

References

  1. WHO (2014) http://www.who.int/mediacentre/factsheets

  2. Myers RM, Carey DE (1967) Concurrent isolation from patient of two arboviruses, Chikungunya and dengue type 2. Science 157:1307–1308

    Article  CAS  PubMed  Google Scholar 

  3. Leroy EM, Nkoghe D, Ollomo B, Nze-Nkogue C, Becquart P, Gerard G, Pourrut X, Charrel R, Moureau G, Ndjoyi-Mbiguino A, De-Lamballerie X (2009) Concurrent chikungunya and dengue virus infections during simultaneous outbreaks, Gabon, 2007. Emerg Infect Dis 15:591–593

    Article  PubMed Central  PubMed  Google Scholar 

  4. Chahar HS, Bharaj P, Dar L, Guleria R, Kabra SK, Broor S (2009) Co-infections with chikungunya virus and dengue virus in Delhi, India. Emerg Infect Dis 5:1077–1080

    Article  Google Scholar 

  5. Kosasih H, de Mast Q, Widjaja S, Sudjana P, Antonjaya U, Ma’roef C, Riswari SF, Porter KR, Burgess TH, Alisjahbana B, van der Ven A, Williams M (2013) Evidence for endemic chikungunya virus infections in Bandung, Indonesia. PLoS Negl Trop Dis 7(10):e2483

    Article  PubMed Central  PubMed  Google Scholar 

  6. Dash PK, Parida MM, Santosh SR, Saxena P, Srivastava A, Neeraja M, Lakshmi V, Rao PV (2008) Development and evaluation of a 1-step duplex reverse transcription polymerase chain reaction for differential diagnosis of chikungunya and dengue infection. Diagn Microbiol Infect Dis 62:52–57

    Article  CAS  PubMed  Google Scholar 

  7. Pongsiri P, Praianantathavorn K, Theamboonlers A, Payungporn S, Poovorawan Y (2012) Multiplex real-time RT-PCR for detecting chikungunya virus and dengue virus. Asian Pac J Trop Med 5(5):342–346

    Article  CAS  PubMed  Google Scholar 

  8. Gurukumar KR, Priyadarshini D, Patil JA, Bhagat A, Singh A, Shah PS, Cecilia D (2009) Development of real time PCR for detection and quantitation of dengue viruses. Virol J 6:10

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  9. 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

    CAS  PubMed Central  PubMed  Google Scholar 

  10. Hasebe F, Parquet MC, Pandey BD, Mathenge EG, Morita K, Balasubramaniam V, Saat Z, Yusop A, Sinniah M, Natkunam S, Igarashi A (2002) Combined detection and genotyping of Chikungunya virus by a specific reverse transcription-polymerase chain reaction. J Med Virol 67:370–374

    Article  CAS  PubMed  Google Scholar 

  11. 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. doi:10.1016/S0166-0934(01)00280-4

    Article  CAS  PubMed  Google Scholar 

  12. Parashar D, Paingankar MS, Kumar S, Gokhale MD, Sudeep AB, Shinde SB, Arankalle VA (2013) Administration of E2 and NS1 siRNAs inhibit chikungunya virus replication in vitro and protects mice infected with the virus. PLoS Negl Trop Dis 7(9):e2405

    Article  PubMed Central  PubMed  Google Scholar 

  13. Mocellin S, Panelli M, Wang E, Rossi CR, Pilati P, Nitti D, Lise M, Marincola FM (2004) IL-10 stimulatory effects on human NK cells explored by gene profile analysis. Genes Immun 5:621–630

    Article  CAS  PubMed  Google Scholar 

  14. Velathanthiri VG, Fernando S, Malavige GN, Peelawatta M, Jayaratne SD, Aaskov J (2006) Comparison of serology, virus isolation and RT-PCR in the diagnosis of dengue viral infections in Sri Lanka. Dengue Bull 30:191

    Google Scholar 

  15. Klungthong C, Gibbons RV, Thaisomboonsuk B, Nisalak A, Kalayanarooj S, Thirawuth V, Nutkumhang N, Mammen MP Jr, Jarman RG (2007) Dengue virus detection using whole blood for reverse transcriptase PCR and virus isolation. J Clin Microbiol 45:2480–2485

    Article  CAS  PubMed Central  PubMed  Google Scholar 

Download references

Acknowledgments

We would like to thank the Director, National Institute of Virology, Pune, for his support, and Dr. Pragya Yadav for testing the KFD and CCHF samples.

Author information

Authors and Affiliations

  1. Dengue/Chikungunya group, National Institute of Virology, Pune, 411001, Maharashtra, India

    D. Cecilia, M. Kakade, K. Alagarasu, J. Patil, A. Salunke, D. Parashar & P. S. Shah

Authors
  1. D. Cecilia
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. M. Kakade
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. K. Alagarasu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. J. Patil
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. A. Salunke
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. D. Parashar
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. P. S. Shah
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to D. Cecilia.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Cecilia, D., Kakade, M., Alagarasu, K. et al. Development of a multiplex real-time RT-PCR assay for simultaneous detection of dengue and chikungunya viruses. Arch Virol 160, 323–327 (2015). https://doi.org/10.1007/s00705-014-2217-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00705-014-2217-x

Keywords

Search

Navigation