Skip to main content

Use of whole blood over plasma enhances the detection of dengue virus RNA: possible utility in dengue vaccine trials

Archives of Virology volume 166, pages 587–591 (2021)Cite this article

Abstract

In the present study, the utility of viral RNA isolated from whole blood over plasma for detection of dengue virus (DENV) was investigated in 80 samples referred for serotyping by DENV serotype-specific one-step real-time RT-PCR. DENV RNA was detected in 71.25% of the whole blood samples compared to 46.25% in the corresponding plasma samples. In secondary infections, DENV RNA was detected in 83.3% of whole blood samples, while it was detected in 40.5% of plasma samples (P = 0.0001). Non-structural protein 1 (NS1) antigen was detected in only 54.8% of the secondary infections. The detection rate of DENV RNA in whole blood is higher than in plasma. We suggest that one-step real-time RT-PCR using RNA from whole blood combined with an NS1 ELISA should be the choice for dengue diagnosis in dengue vaccine trials.

This is a preview of subscription content, access via your institution.

Access options

Buy single article

Instant access to the full article PDF.

43,34 €

Price includes VAT (Finland)
Tax calculation will be finalised during checkout.

References

  1. Muller DA, Depelsenaire ACI, Young PR (2017) Clinical and laboratory diagnosis of dengue virus infection. J Infect Dis 215:S89–S95. https://doi.org/10.1093/infdis/jiw649

    CAS  Article  PubMed  Google Scholar 

  2. Andries AC, Duong V, Ly S et al (2015) Value of routine dengue diagnostic tests in urine and saliva specimens. PLoS Negl Trop Dis 9(9):e0004100. https://doi.org/10.1371/journal.pntd.0004100

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  3. World Health Organization null (2017) Dengue vaccine: WHO position paper, July 2016—recommendations. Vaccine 35:1200–1201. https://doi.org/10.1016/j.vaccine.2016.10.070

    Article  Google Scholar 

  4. World Health Organization (2012) Guidelines on the quality, safety and efficacy of dengue tetravalent vaccines (live, attenuated). www.who.int/biologicals/vaccines/Dengue_DB_5_April_2012_clean.pdf. Accessed 1 Oct 2020.

  5. Kakade MB, Shrivastava N, Patil JA et al (2020) Clinical evaluation of an in-house-developed real-time RT-PCR assay for serotyping of dengue virus. Arch Virol 165:2311–2315. https://doi.org/10.1007/s00705-020-04725-0

    CAS  Article  PubMed  Google Scholar 

  6. Kulkarni R, Modak M, Gosavi M et al (2020) Comparative assessment of commercial enzyme-linked immunosorbent assay and rapid diagnostic tests used for dengue diagnosis in India. Indian J Med Res 151:71–78. https://doi.org/10.4103/ijmr.IJMR_613_18

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  7. Klungthong C, Gibbons RV, Thaisomboonsuk B et al (2007) Dengue virus detection using whole blood for reverse transcriptase PCR and virus isolation. J Clin Microbiol 45:2480–2485. https://doi.org/10.1128/JCM.00305-07

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  8. Kar M, Nisheetha A, Kumar A et al (2019) Isolation and molecular characterization of dengue virus clinical isolates from pediatric patients in New Delhi. Int J Infect Dis 84S:S25–S33. https://doi.org/10.1016/j.ijid.2018.12.003

    CAS  Article  PubMed  Google Scholar 

  9. De Paula SO, Lopes da Fonseca BA (2002) Optimizing dengue diagnosis by RT-PCR in IgM-positive samples: comparison of whole blood, buffy-coat and serum as clinical samples. J Virol Methods 102:113–117. https://doi.org/10.1016/s0166-0934(02)00005-8

    Article  PubMed  Google Scholar 

  10. De Paula SO, de Melo LC, Torres MP et al (2004) One-step RT-PCR protocols improve the rate of dengue diagnosis compared to two-step RT-PCR approaches. J Clin Virol 30:297–301. https://doi.org/10.1016/j.jcv.2003.11.004

    CAS  Article  PubMed  Google Scholar 

  11. Cholet F, Ijaz UZ, Smith CJ (2019) Differential ratio amplicons (Ramp) for the evaluation of RNA integrity extracted from complex environmental samples. Environ Microbiol 21:827–844. https://doi.org/10.1111/1462-2920.14516

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  12. Añez G, Heisey DAR, Chancey C et al (2016) Distribution of dengue virus types 1 and 4 in blood components from infected blood donors from Puerto Rico. PLoS Negl Trop Dis 10:e0004445. https://doi.org/10.1371/journal.pntd.0004445

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  13. Shan X, Wang X, Yuan Q et al (2015) Evaluation of the diagnostic accuracy of nonstructural protein 1 Ag-based tests for dengue virus in Asian population: a meta-analysis. BMC Infect Dis 15:360. https://doi.org/10.1186/s12879-015-1088-4

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  14. World Health Organization (2009) Dengue guidelines for diagnosis, treatment, prevention and control: new edition. World Health Organization. https://apps.who.int/iris/handle/10665/44188. Accessed 27 July 2020

  15. Plennevaux E, Sabchareon A, Limkittikul K et al (2016) Detection of dengue cases by serological testing in a dengue vaccine efficacy trial: utility for efficacy evaluation and impact of future vaccine introduction. Vaccine 34:2707–2712. https://doi.org/10.1016/j.vaccine.2016.04.028

    CAS  Article  PubMed  Google Scholar 

  16. Plennevaux E, Moureau A, Arredondo-García JL et al (2018) Impact of dengue vaccination on serological diagnosis: insights from phase III dengue vaccine efficacy trials. Clin Infect Dis 66:1164–1172. https://doi.org/10.1093/cid/cix966

    CAS  Article  PubMed  Google Scholar 

  17. Alagarasu K (2016) Introducing dengue vaccine: implications for diagnosis in dengue vaccinated subjects. Vaccine 34:2759–2761. https://doi.org/10.1016/j.vaccine.2016.04.070

    Article  PubMed  Google Scholar 

  18. Guo C, Zhou Z, Wen Z et al (2017) Global epidemiology of dengue outbreaks in 1990–2015: a systematic review and meta-analysis. Front Cell Infect Microbiol 7:317. https://doi.org/10.3389/fcimb.2017.00317

    Article  PubMed  PubMed Central  Google Scholar 

Download references

Author information

Affiliations

  1. Dengue and Chikungunya Group, ICMR-National Institute of Virology, Pune, Maharashtra, 411001, India

    K. Alagarasu, M. B. Kakade, R. V. Bachal, M. Bote, D. Parashar & P. S. Shah

Authors
  1. K. Alagarasu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. M. B. Kakade
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. R. V. Bachal
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. M. Bote
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. D. Parashar
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. P. S. Shah
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

KA, DP and PSS conceived and designed the study. MBK, RVB, and MB performed the study. KA did the statistical analysis and wrote the manuscript. DP critically reviewed the manuscript, and all of the authors approved the content.

Corresponding author

Correspondence to K. Alagarasu.

Ethics declarations

Conflict of interest

None declared.

Ethical approval

Approved by the Institutional Ethics committee of ICMR-NIV [NIV/IEC/2018/D-8 and NIV/IEC/Jan/2020/D-6].

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Handling Editor: William G Dundon.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary file1 (PPTX 171 KB)

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Alagarasu, K., Kakade, M.B., Bachal, R.V. et al. Use of whole blood over plasma enhances the detection of dengue virus RNA: possible utility in dengue vaccine trials. Arch Virol 166, 587–591 (2021). https://doi.org/10.1007/s00705-020-04892-0

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00705-020-04892-0