Display options
Cite
Belachew M, Wolde M, Nega D, et al. Evaluating performance of multiplex real time PCR for the diagnosis of malaria at elimination targeted low transmission settings of Ethiopia. Malar J. 2022;21(1):9doi: 10.1186/s12936-021-04029-x.
Belachew, M., Wolde, M., Nega, D., Gidey, B., Negash, L., Assefa, A., Tasew, G., Woyessa, A., & Abera, A. (2022). Evaluating performance of multiplex real time PCR for the diagnosis of malaria at elimination targeted low transmission settings of Ethiopia. Malaria journal, 21(1), 9. https://doi.org/10.1186/s12936-021-04029-x
Belachew, Mahlet, et al. "Evaluating performance of multiplex real time PCR for the diagnosis of malaria at elimination targeted low transmission settings of Ethiopia." Malaria journal vol. 21,1 (2022): 9. doi: https://doi.org/10.1186/s12936-021-04029-x
Belachew M, Wolde M, Nega D, Gidey B, Negash L, Assefa A, Tasew G, Woyessa A, Abera A. Evaluating performance of multiplex real time PCR for the diagnosis of malaria at elimination targeted low transmission settings of Ethiopia. Malar J. 2022 Jan 06;21(1):9. doi: 10.1186/s12936-021-04029-x. PMID: 34986840; PMCID: PMC8734331.
Copy Download .nbib Format: NLM
Share it on

Malar J. 2022 Jan 06;21(1):9. doi: 10.1186/s12936-021-04029-x.

Evaluating performance of multiplex real time PCR for the diagnosis of malaria at elimination targeted low transmission settings of Ethiopia.

Malaria journal

Mahlet Belachew, Mistire Wolde, Desalegn Nega, Bokretsion Gidey, Legessie Negash, Ashenafi Assefa, Geremew Tasew, Adugna Woyessa, Adugna Abera

Affiliations

  1. Department of Medical Laboratory Sciences, College of Health Sciences, Addis Ababa University, Addis Ababa, Ethiopia.
  2. Malaria and Neglected Tropical Diseases Research Team, Ethiopian Public Health Institute, P.O. Box: 1242, Addis Ababa, Ethiopia.
  3. Malaria and Neglected Tropical Diseases Research Team, Ethiopian Public Health Institute, P.O. Box: 1242, Addis Ababa, Ethiopia. [email protected].

PMID: 34986840 PMCID: PMC8734331 DOI: 10.1186/s12936-021-04029-x

Abstract

BACKGROUND: Malaria incidence has declined in Ethiopia in the past 10 years. Current malaria diagnostic tests, including light microscopy and rapid antigen-detecting diagnostic tests (RDTs) cannot reliably detect low-density infections. Studies have shown that nucleic acid amplification tests are highly sensitive and specific in detecting malaria infection. This study took place with the aim of evaluating the performance of multiplex real time PCR for the diagnosis of malaria using patient samples collected from health facilities located at malaria elimination targeted low transmission settings in Ethiopia.

METHODS: A health facility-based, cross-sectional survey was conducted in selected malaria sentinel sites. Malaria-suspected febrile outpatients referred to laboratory for malaria testing between December 2019 and March 2020 was enrolled into this study. Sociodemographic information and capillary blood samples were collected from the study participants and tested at spot with RDTs. Additionally, five circles of dry blood spot (DBS) samples on Whatman filter paper and thick and thin smear were prepared for molecular testing and microscopic examination, respectively. Multiplex real time PCR assay was performed at Ethiopian Public Health Institute (EPHI) malaria laboratory. The performance of multiplex real time PCR assay, microscopy and RDT for the diagnosis of malaria was compared and evaluated against each other.

RESULTS: Out of 271 blood samples, multiplex real time PCR identified 69 malaria cases as Plasmodium falciparum infection, 16 as Plasmodium vivax and 3 as mixed infections. Of the total samples, light microscopy detected 33 as P. falciparum, 18 as P. vivax, and RDT detected 43 as P. falciparum, 17 as P. vivax, and one mixed infection. Using light microscopy as reference test, the sensitivity and specificity of multiplex real time PCR were 100% (95% CI (93-100)) and 83.2% (95% CI (77.6-87.9)), respectively. Using multiplex real time PCR as a reference, light microscopy and RDT had sensitivity of 58% (95% CI 46.9-68.4) and 67% (95% CI 56.2-76.7); and 100% (95% CI 98-100) and 98.9% (95% CI 96-99.9), respectively. Substantial level of agreement was reported between microscopy and multiplex real time PCR results with kappa value of 0.65.

CONCLUSIONS: Multiplex real-time PCR had an advanced performance in parasite detection and species identification on febrile patients' samples than did microscopy and RDT in low malaria transmission settings. It is highly sensitive malaria diagnostic method that can be used in malaria elimination programme, particularly for community based epidemiological samples. Although microscopy and RDT had reduced performance when compared to multiplex real time PCR, still had an acceptable performance in diagnosis of malaria cases on patient samples at clinical facilities.

© 2021. The Author(s).

Keywords: Diagnostic performance; Malaria elimination; Multiplex real time PCR

References

  1. Malar J. 2017 Aug 10;16(1):328 - PubMed
  2. Malar J. 2016 Feb 16;15:88 - PubMed
  3. J Clin Microbiol. 2004 Dec;42(12):5636-43 - PubMed
  4. Emerg Infect Dis. 2015 Oct;21(10):1853-7 - PubMed
  5. Malar J. 2012 Jul 20;11:234 - PubMed
  6. Clin Microbiol Infect. 2011 Mar;17(3):469-75 - PubMed
  7. Eur J Clin Microbiol Infect Dis. 2018 Dec;37(12):2323-2329 - PubMed
  8. Am J Trop Med Hyg. 2002 Jun;66(6):641-8 - PubMed
  9. Eur J Clin Microbiol Infect Dis. 2017 Apr;36(4):671-675 - PubMed
  10. Malar J. 2011 Jun 26;10:175 - PubMed
  11. Malar J. 2018 Jan 12;17(1):26 - PubMed
  12. Malar J. 2014 Feb 06;13:48 - PubMed
  13. Cold Spring Harb Perspect Med. 2017 Sep 1;7(9): - PubMed
  14. Cell. 2016 Oct 20;167(3):610-624 - PubMed
  15. Parasite Epidemiol Control. 2019 Apr 25;6:e00107 - PubMed
  16. Int J Environ Res Public Health. 2018 Feb 03;15(2): - PubMed
  17. Infect Dis Poverty. 2018 Nov 5;7(1):103 - PubMed
  18. Am J Trop Med Hyg. 2015 Jul;93(1):181-5 - PubMed
  19. BMC Infect Dis. 2017 Jun 6;17(1):399 - PubMed
  20. J Infect Dis. 2015 May 1;211(9):1476-83 - PubMed
  21. EBioMedicine. 2020 May;55:102757 - PubMed
  22. Adv Clin Chem. 2017;80:155-192 - PubMed
  23. Mem Inst Oswaldo Cruz. 2011 Sep;106(6):691-700 - PubMed
  24. Curr Opin Infect Dis. 2015 Oct;28(5):446-54 - PubMed
  25. PLoS One. 2019 Jul 5;14(7):e0218982 - PubMed
  26. Malar J. 2013 Oct 03;12:352 - PubMed
  27. Malar J. 2014 Oct 18;13:411 - PubMed
  28. BMC Public Health. 2020 Mar 12;20(1):320 - PubMed
  29. Malar J. 2009 Dec 09;8:284 - PubMed

Publication Types