Detection and differentiation of dengue virus serotypes by one-step multiplex reverse transcription PCR assays / Detecção e diferenciação de sorotipos do vírus da dengue por ensaios de PCR com transcrição reversa multiplexada em uma etapa

Authors

  • Mariana Maryelle Ferreira de Sousa
  • Bruna de Oliveira de Melo
  • Antonia Khaynnam Silva Costa
  • Jadna Patricia Pinheiro Nunes
  • Silvio Gomes Monteiro
  • Ana Cláudia Pinho de Carvalho
  • Conceição de Maria Fernandes da Silva Pinto
  • Lécia Maria Sousa Santos Cosme
  • Rosimary de Jesus Gomes Turri
  • Mauro Martins Teixeira
  • Maria Rosa Quaresma Bomfim

DOI:

https://doi.org/10.34117/bjdv6n1-016

Keywords:

Molecular diagnostics, dengue infections, MAC-ELISA, One-step multiplex RT-PCR assay.

Abstract

Background: Dengue infections are a severe public health problem in Brazil. The Ministry of Health recommends an immunosorbent assay (ELISA) for the capture of IgM (MAC-ELISA) to diagnose dengue. However, it detects antibodies that cross-react with other flaviviruses and requires confirmation in reference laboratories. Methods: One-step multiplex RT-PCR assay was used to amplify RNA of 197 serum from patients with clinical suspicion of dengue infection. The samples had been screened with the IgM ELISA kit in the Central Public Health Laboratory of the State of Maranhão. Results: Of the 197 samples evaluated by IgM ELISA, 135 were positive; of these, 96 (71.1%) were from patients in the acute phase of the infection. The one-step multiplex RT-PCR detected viral RNA in 88 (91.7%) of this serum. Among the 62-negative serum by ELISA, 29 samples (46.8%) were amplified using the molecular method. Conclusions: One-step multiplex RT-PCR was sensitive in the detection of viral particles from the first day until the sixth day after the onset of the feverish period. Moreover, it was specific and 100% reproducible. Based on these results, we recommend the use of this molecular assay to diagnose and differentiate the DENV serotypes in the acute phase of the disease.

References

Normile D. Surprising new dengue virus throws a spanner in disease control efforts. Science. 2013;342(6157):415.

Mustafa MS, Rasotgi V, Jain S, Gupta V. Discovery of fifth serotype of dengue virus (DENV-5): A new public health dilemma in dengue control. Med J Armed Forces India. 2015;71(1):67-70.

Monteiro VVS, Navegantes-Lima KC, de Lemos AB, da Silva GL, de Souza Gomes R, Reis JF, Rodrigues Junior LC, da Silva OS, Romão PRT, Monteiro MC. Aedes-Chikungunya Virus Interaction: Key Role of Vector Midguts Microbiota and Its Saliva in the Host Infection. Front Microbiol. 2019 Apr 9; 10:492. doi: 10.3389/fmicb.2019.00492. PMID: 31024463; PMCID: PMC6467098.

Murray NE, Quam MB, Wilder-Smith A. Epidemiology of dengue: past, present and future prospects. Clin Epidemiol. 2013; 5:299–309.

Liang G, Gao X, Gould, EA. Factors responsible for the emergence of arboviruses; strategies, challenges and limitations for their control. Emerg Microb Infect. 2015; 4(3):e18.

Qinlong J, Wang M. Dengue epidemiology. Global Health Journal June 30, 2019, Volume 3, Issue 2

Guzman MG, Gubler DJ, Izquierdo A, Martinez E, Haltead SB. Dengue infection. Nat Rev Dis Primers 2, 16055 (2016) doi:10.1038/nrdp.2016.55.

Khetarpal N, Khanna I. Dengue Fever: Causes, Complications, and Vaccine Strategies. J Immunol Res vol. 2016, Article ID 6803098, 14 pages, 2016. https://doi.org/10.1155/2016/6803098.

World Health Organization (WHO). Dengue fact sheet. [Internet]. Regional Office for South-East Asia. Dengue fact sheet. [cited 2019 october]. Available in:http://www.searo.who.int/entity/vector_borne_tropical_diseases/data/data_factsheet/en/

World Health Organization (WHO). Dengue and severe dengue. Geneva. Cogan JE; 4 November 2019. Dengue and severe dengue. [cited 2019 November 19]. Available in:https://www.who.int/news-room/fact-sheets/detail/dengue-and-severe-dengue.

Ministério da Saúde (MS). Secretaria de Vigilância em Saúde. Sistema de Informação de Agravos de Notificação (SINAN). [cited 2019 october 31]. Available in:http://portalarquivos2.saude.gov.br/images/pdf/2017/fevereiro/10/obitos-ate-2016.pdf.

Ministério da Saúde (MS). Secretaria de Vigilância em Saúde. Boletins Epidemiológicos sobre a situação epidemiológica dos casos de Dengue. [cited 2019 May 6]. Available in: http://www.saude.gov.br/boletins-epidemiologicos.

Pawar R, Patravale V. Dengue Diagnosis: Challenges and Opportunities. Immunochem Immunopathol. 2015;2015(1):1-10.

Muller DA, Depelsenaire AC, Young PR. Clinical and Laboratory Diagnosis of Dengue Virus Infection. J Infect Dis. 2017;215:S89-S95.

Srinivas V, Srinivas VR. Dengue Fever: A Review Article. J Evol Med Dent Sci. 2015;4(29):5048-5058.

Hu D, Di B, Ding X, Wang Y, Chen Y, Pan Y, et al. Kinetics of non-structural protein 1, IgM and IgG antibodies in dengue type 1 primary infection. Virol J. 2011; 8:47.

Nascimento EJM, Huleatt JW, Cordeiro MT, Castanha PMS, George JK, Grebe E, Welte A, Brown M, Burke DS, Marques ETA. Development of antibody biomarkers of long term and recent dengue virus infections. J Virol Methods, 257 (2018), pp. 62-68.

Mardekian SK, Roberts AL. Diagnostic Options and Challenges for dengue and Chikungunya Viruses. Biomed Res Int. 2015; 2015:834371. doi: 10.1155/2015/834371. Epub 2015 Oct 5. PMID: 26509163; PMCID: PMC4609775.

Lanciotti RS, Calisher CH, Guble DJ, Chang GJ, Vorndam AV. Rapid detection and typing of Dengue viruses from clinical samples by using reverse transcriptase-polymerase chain reaction. J Clin Microbiol. 1992;30(3):545-51.

Nunes JPP, Melo BO, Monteiro SG, Almeida VSS, Monteiro AS, Cosme LMSS, Pinto CMFS, Luena MSS, Turri RJG, Mendes MFC, Bomfim MRQ. One-Step reverse transcriptase PCR for detection of arboviruses in serum samples of patients assisted in Basic health Units in the State of Maranhão, Brazil.Braz. J. of Develop., Curitiba, v. 5, n. 9, p. 16620-16644 sep. 2019.

Sambrook J, Fritsch EF, Maniatis T. Molecular Cloning: a laboratory manual. 2nd ed. N.Y., Cold Spring Harbor Laboratory, Cold Spring Harbor Laboratory Press, 1659 p.

IBM SPSS Statistics for Windows, Version 20.0. Armonk, NY: IBM Corp. 2011.

Bhatnagar J, Blau DM, Shieh WJ, Paddock CD, Drew C, Liu L, et al. Molecular detection and typing of dengue viruses from archived tissues of fatal cases by rt-PCR and sequencing: diagnostic and epidemiologic implications. Am J Trop Med Hyg. 2013;86(2):335-40.

Bandyopadhyay B, Bhattacharyya I, Adhikary S, Konar J, Dawar N, Sarkar J, et al. A Comprehensive Study on the 2012 Dengue Fever Outbreak in Kolkata, India. Virol. 2013; 2013:5.

Ferraz FO, Bomfim MRQ, Totola AH, Ávila TV, Cisalpino D, Pessanha JEM, et al. Evaluation of laboratory tests for dengue diagnosis in clinical specimens from consecutive patients with suspected dengue in Belo Horizonte, Brazil. J Clini Virol. 2013;58(1):41-46.

Yong YK, Thayan R, Chong HT, Tan CT, Sekaran SD. Rapid detection and serotyping of dengue virus by multiplex RT-PCR and real-time SYBR green RT-PCR. Singapore Med J. 2007;48(7):662-8.

Tontulawat P, Pongsiri P, Thongmee C, Theamboonlers A, Kamolvarin N, Poovorawan Y. Evaluation of rapid immunochromatographic NS1 test, anti-dengue IgM test, semi-nested PCR and IgM ELISA for detection of dengue virus. Southeast Asian J Trop Med Public Health. 2011;42(3):570–8.

Chien LJ, Liao TL, Shu PY, Huang JH, Gubler DJ, Chang GJ. Development of real-time reverse transcriptase PCR assays to detect and serotype dengue viruses. J Clin Microbiol. 2009;44(4):1295-304.

Ahmed NH, Broor S. Comparison of NS1 antigen detection ELISA, real time RT-PCR and virus isolation for rapid diagnosis of dengue infection in acute phase. J Vector Borne Dis. 2014;51(3):194–9.

Alm E, Lindegren G, Falk KI, Lagerqvist N. One-step real-time RT-PCR assays for serotyping dengue virus in clinical samples. BMC Infect Dis. 2015; 15:493.

Simmons M, Myers T, Guevara C, Jungkind D, Williams M, Houng HS. Development and Validation of a Quantitative, One-Step, Multiplex, Real-Time Reverse Transcriptase PCR Assay for Detection of Dengue and Chikungunya Viruses. J Clin Microbiol. 2016;54(7):1766-1773

Mun MJ, YongBae J, HyuckKim J, BokKim S, Lee I, IlKim J, et al. One-step multiplex real-time RT-PCR for detection and typing of dengue vírus. Molec Cell Prob. 2019;43:86-91.

Waggoner JJ, Abeynayake J, Sahoo MK, Gresh L, Tellez Y, Gonzalez K, et al. Single-Reaction, Multiplex, Real-Time RT-PCR for the Detection, Quantitation, and Serotyping of Dengue Viruses. PLoS Negl Trop Dis. 2013;7:e2116

Waggoner JJ, Gresh L, Mohamed-Hadley A, Ballesteros G, Davila MJ, Tellez Y, et al. Single-Reaction Multiplex Reverse Transcription PCR for Detection of Zika, Chikungunya, and Dengue Viruses. Emerg Infect Dis. 2016;22(7):1295–1297.

Hajia M. Limitations of Different PCR Protocols Used in Diagnostic Laboratories: A Short Review, Modern Med Labor J. 2017;1(1):1-6.

Branco MS, Sousa DM, Sousa DM, Monteiro JD, Costa DM, Monteiro JD, et al. Dengue in the State of Rio Grande do Norte, Brazil, 2010–2012. Trop Med Intern Health. 2015; 20:1707-1710.

Costa CAD, Façanha GP. Dengue virus serotypes in children of Manaus, State of Amazonas, 2008. Rev Soc Bras Med Trop. 2011;44(2): 249-251.

Pinho de Carvalho AC, Couto Portela F, Azevedo Feitosa Ferro T, Quaresma Bomfim MR. EPIDEMIOLOGY OF DENGUE VIRUS FROM 2002-2012 IN SAO LUIS, MARANHAO STATE, BRAZIL. J Trop Pathol. 2016;45(3):243-255

Assunção ML, Aguilar AMM. Perfil Clínico-epidemiológico da dengue no Município de Juscimeira-MT. Rev. Epidemiol Control Infec, 2015;4(4):249-253.

Gubler DJ. Dengue, Urbanization and Globalization: The Unholy Trinity of the 21(st) Century. Trop Med Health. 2011; 39(4 Suppl):3-11.

Amaku M, Azevedo F, Burattini MN, Coelho GE, Coutinho FAB, Greenhalgh D, et al. Magnitude and frequency variations of vector-borne infection outbreaks using the Ross–Macdonald model: explaining and predicting outbreaks of dengue fever. Epidemiol. Infect. 2016;144(16):3435-3450.

Akter R, Naish S, Hu W, Tong S. Socio-demographic, ecological factors and dengue infection trends in Australia. PLoS One. 2017;12(10):e0185551.

Simmonds P, Becher P, Bukh J, Gould EA, Meyers G, Monath T, et al. ICTV Virus Taxonomy Profile: Flaviviridae. J Gen Virol. 2017;98(1):2–3.

Reller LB, Weinstein MP, Petti CA, Detection and Identification of Microorganisms by Gene Amplification and Sequencing, Clin Infect Dis. 2007;44(8):1108–1114.

Stadhouders R, Pas S, Anber J, Voermans J, Mes THM, Schutten M. The Effect of Primer-Template Mismatches on the Detection and Quantification of Nucleic Acids Using the 5? Nuclease Assay. J Molec Diagn. 2010;12(1):109-17.

Clyde K, Kyle JL, Harris E. Recent advances in deciphering viral and host determinants of dengue virus replication and pathogenesis. J Virol. 2006;80 (23):11418–11431.

Rastogi M, Sharma N, Singh SK. Flavivirus NS1: a multifaceted enigmatic viral protein. Virol J. 2016; 13:131.

World Health Organization (WHO). Dengue guidelines for diagnosis, treatment, prevention and control. Chapter 4. Laboratory diagnosis and diagnostic tests. Geneva: WHO; 2009.91-106p.

Kim JH, Chong CK, Sinnadurai J, Song HO, Park H. Clinical diagnosis of early dengue infection by novel one-step multiplex real-time RT-PCR targeting NS1 gene. J Clin Virol. 2015; 65:11-9.

Published

2020-01-03

How to Cite

Sousa, M. M. F. de, Melo, B. de O. de, Costa, A. K. S., Nunes, J. P. P., Monteiro, S. G., Carvalho, A. C. P. de, Pinto, C. de M. F. da S., Cosme, L. M. S. S., Turri, R. de J. G., Teixeira, M. M., & Bomfim, M. R. Q. (2020). Detection and differentiation of dengue virus serotypes by one-step multiplex reverse transcription PCR assays / Detecção e diferenciação de sorotipos do vírus da dengue por ensaios de PCR com transcrição reversa multiplexada em uma etapa. Brazilian Journal of Development, 6(1), 227–246. https://doi.org/10.34117/bjdv6n1-016

Issue

Section

Original Papers