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Published in: Malaria Journal 1/2019

Open Access 01-12-2019 | Malaria | Research

Field evaluation of malaria malachite green loop-mediated isothermal amplification in health posts in Roraima state, Brazil

Authors: Heather M. Kudyba, Jaime Louzada, Dragan Ljolje, Karl A. Kudyba, Vasant Muralidharan, Joseli Oliveira-Ferreira, Naomi W. Lucchi

Published in: Malaria Journal | Issue 1/2019

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Abstract

Background

Microscopic detection of malaria parasites is the standard method for clinical diagnosis of malaria in Brazil. However, malaria epidemiological surveillance studies specifically aimed at the detection of low-density infection and asymptomatic cases will require more sensitive and field-usable tools. The diagnostic accuracy of the colorimetric malachite green, loop-mediated, isothermal amplification (MG-LAMP) assay was evaluated in remote health posts in Roraima state, Brazil.

Methods

Study participants were prospectively enrolled from health posts (healthcare-seeking patients) and from nearby villages (healthy participants) in three different study sites. The MG-LAMP assay and microscopy were performed in the health posts. Two independent readers scored the MG-LAMP tests as positive (blue/green) or negative (clear). Sensitivity and specificity of local microscopy and MG-LAMP were calculated using results of PET-PCR as a reference.

Results

A total of 91 participants were enrolled. There was 100% agreement between the two MG-LAMP readers (Kappa = 1). The overall sensitivity and specificity of MG-LAMP were 90.0% (95% confidence interval (CI) 76.34–97.21%) and 94% (95% CI 83.76–98.77%), respectively. The sensitivity and specificity of local microscopy were 83% (95% CI 67.22–92.66%) and 100% (95% CI 93.02–100.00%), respectively. PET-PCR detected six mixed infections (infection with both Plasmodium falciparum and Plasmodium vivax); two of these were also detected by MG-LAMP and one by microscopy. Microscopy did not detect any Plasmodium infection in the 26 healthy participants; MG-LAMP detected Plasmodium in five of these and PET-PCR assay detected infection in three. Overall, performing the MG-LAMP in this setting did not present any particular challenges.

Conclusion

MG-LAMP is a sensitive and specific assay that may be useful for the detection of malaria parasites in remote healthcare settings. These findings suggest that it is possible to implement simple molecular tests in facilities with limited resources.
Literature
1.
go back to reference WHO. World malaria report. Geneva: World Health Organization; 2018. WHO. World malaria report. Geneva: World Health Organization; 2018.
2.
go back to reference PAHO. Epidemiological alert: increase of malaria in the Americas. Washington: Pan American Health Organization; 2018. PAHO. Epidemiological alert: increase of malaria in the Americas. Washington: Pan American Health Organization; 2018.
3.
go back to reference Wongsrichanalai C, Barcus MJ, Muth S, Sutamihardja A, Wernsdorfer WH. A review of malaria diagnostic tools: microscopy and rapid diagnostic test (RDT). Am J Trop Med Hyg. 2007;77:119–27.CrossRef Wongsrichanalai C, Barcus MJ, Muth S, Sutamihardja A, Wernsdorfer WH. A review of malaria diagnostic tools: microscopy and rapid diagnostic test (RDT). Am J Trop Med Hyg. 2007;77:119–27.CrossRef
4.
go back to reference Tajebe A, Magoma G, Aemero M, Kimani F. Detection of mixed infection level of Plasmodium falciparum and Plasmodium vivax by SYBR Green I-based real-time PCR in North Gondar, north-west Ethiopia. Malar J. 2014;3:411.CrossRef Tajebe A, Magoma G, Aemero M, Kimani F. Detection of mixed infection level of Plasmodium falciparum and Plasmodium vivax by SYBR Green I-based real-time PCR in North Gondar, north-west Ethiopia. Malar J. 2014;3:411.CrossRef
5.
go back to reference Ehtesham R, Fazaeli A, Raeisi A, Keshavarz H, Heidari A. Detection of mixed-species infections of Plasmodium falciparum and Plasmodium vivax by nested PCR and rapid diagnostic tests in southeastern Iran. Am J Trop Med Hyg. 2015;93:181–5.CrossRef Ehtesham R, Fazaeli A, Raeisi A, Keshavarz H, Heidari A. Detection of mixed-species infections of Plasmodium falciparum and Plasmodium vivax by nested PCR and rapid diagnostic tests in southeastern Iran. Am J Trop Med Hyg. 2015;93:181–5.CrossRef
6.
go back to reference Krishna S, Bharti PK, Chandel HS, Ahmad A, Kumar R, Singh PP, et al. Detection of mixed infections with Plasmodium spp. by PCR, India, 2014. Emerg Infect Dis. 2015;21:1853–7.CrossRef Krishna S, Bharti PK, Chandel HS, Ahmad A, Kumar R, Singh PP, et al. Detection of mixed infections with Plasmodium spp. by PCR, India, 2014. Emerg Infect Dis. 2015;21:1853–7.CrossRef
7.
go back to reference Roth JM, Korevaar DA, Leeflang MM, Mens PF. Molecular malaria diagnostics: a systematic review and meta-analysis. Crit Rev Clin Lab Sci. 2016;53:87–105.CrossRef Roth JM, Korevaar DA, Leeflang MM, Mens PF. Molecular malaria diagnostics: a systematic review and meta-analysis. Crit Rev Clin Lab Sci. 2016;53:87–105.CrossRef
8.
go back to reference Okell LC, Ghani AC, Lyons E, Drakeley CJ. Submicroscopic infection in Plasmodium falciparum-endemic populations: a systematic review and meta-analysis. J Infect Dis. 2009;200:1509–17.CrossRef Okell LC, Ghani AC, Lyons E, Drakeley CJ. Submicroscopic infection in Plasmodium falciparum-endemic populations: a systematic review and meta-analysis. J Infect Dis. 2009;200:1509–17.CrossRef
9.
go back to reference WHO. A framework for malaria elimination. Geneva: World Health Organization; 2017. WHO. A framework for malaria elimination. Geneva: World Health Organization; 2017.
11.
go back to reference Cook J, Aydin-Schmidt B, Gonzalez IJ, Bell D, Edlund E, Nassor MH, et al. Loop-mediated isothermal amplification (LAMP) for point-of-care detection of asymptomatic low-density malaria parasite carriers in Zanzibar. Malar J. 2015;14:43.CrossRef Cook J, Aydin-Schmidt B, Gonzalez IJ, Bell D, Edlund E, Nassor MH, et al. Loop-mediated isothermal amplification (LAMP) for point-of-care detection of asymptomatic low-density malaria parasite carriers in Zanzibar. Malar J. 2015;14:43.CrossRef
12.
go back to reference Hopkins H, Gonzalez IJ, Polley SD, Angutoko P, Ategeka J, Asiimwe C, et al. Highly sensitive detection of malaria parasitemia in a malaria-endemic setting: performance of a new loop-mediated isothermal amplification kit in a remote clinic in Uganda. J Infect Dis. 2013;208:645–52.CrossRef Hopkins H, Gonzalez IJ, Polley SD, Angutoko P, Ategeka J, Asiimwe C, et al. Highly sensitive detection of malaria parasitemia in a malaria-endemic setting: performance of a new loop-mediated isothermal amplification kit in a remote clinic in Uganda. J Infect Dis. 2013;208:645–52.CrossRef
13.
go back to reference Lucchi NW, Demas A, Narayanan J, Sumari D, Kabanywanyi A, Kachur SP, et al. Real-time fluorescence loop mediated isothermal amplification for the diagnosis of malaria. PLoS ONE. 2010;5:e13733.CrossRef Lucchi NW, Demas A, Narayanan J, Sumari D, Kabanywanyi A, Kachur SP, et al. Real-time fluorescence loop mediated isothermal amplification for the diagnosis of malaria. PLoS ONE. 2010;5:e13733.CrossRef
14.
go back to reference Polley SD, Mori Y, Watson J, Perkins MD, Gonzalez IJ, Notomi T, et al. Mitochondrial DNA targets increase sensitivity of malaria detection using loop-mediated isothermal amplification. J Clin Microbiol. 2010;48:2866–71.CrossRef Polley SD, Mori Y, Watson J, Perkins MD, Gonzalez IJ, Notomi T, et al. Mitochondrial DNA targets increase sensitivity of malaria detection using loop-mediated isothermal amplification. J Clin Microbiol. 2010;48:2866–71.CrossRef
15.
go back to reference Oriero EC, Jacobs J, Van Geertruyden JP, Nwakanma D, D’Alessandro U. Molecular-based isothermal tests for field diagnosis of malaria and their potential contribution to malaria elimination. J Antimicrob Chemother. 2015;70:2–13.CrossRef Oriero EC, Jacobs J, Van Geertruyden JP, Nwakanma D, D’Alessandro U. Molecular-based isothermal tests for field diagnosis of malaria and their potential contribution to malaria elimination. J Antimicrob Chemother. 2015;70:2–13.CrossRef
16.
go back to reference Lucchi NW, Ndiaye D, Britton S, Udhayakumar V. Expanding the malaria molecular diagnostic options: opportunities and challenges for loop-mediated isothermal amplification tests for malaria control and elimination. Expert Rev Mol Diagn. 2018;18:195–203.CrossRef Lucchi NW, Ndiaye D, Britton S, Udhayakumar V. Expanding the malaria molecular diagnostic options: opportunities and challenges for loop-mediated isothermal amplification tests for malaria control and elimination. Expert Rev Mol Diagn. 2018;18:195–203.CrossRef
17.
go back to reference Lucchi NW, Ljolje D, Silva-Flannery L, Udhayakumar V. Use of malachite green-loop mediated isothermal amplification for detection of Plasmodium spp. parasites. PLoS ONE. 2016;11:e0151437.CrossRef Lucchi NW, Ljolje D, Silva-Flannery L, Udhayakumar V. Use of malachite green-loop mediated isothermal amplification for detection of Plasmodium spp. parasites. PLoS ONE. 2016;11:e0151437.CrossRef
18.
go back to reference Britton S, Cheng Q, Grigg MJ, William T, Anstey NM, McCarthy JS. A sensitive, colorimetric, high-throughput loop-mediated isothermal amplification assay for the detection of Plasmodium knowlesi. Am J Trop Med Hyg. 2016;95:120–2.CrossRef Britton S, Cheng Q, Grigg MJ, William T, Anstey NM, McCarthy JS. A sensitive, colorimetric, high-throughput loop-mediated isothermal amplification assay for the detection of Plasmodium knowlesi. Am J Trop Med Hyg. 2016;95:120–2.CrossRef
19.
go back to reference Britton S, Cheng Q, Sutherland CJ, McCarthy JS. A simple, high-throughput, colourimetric, field applicable loop-mediated isothermal amplification (HtLAMP) assay for malaria elimination. Malar J. 2015;14:335.CrossRef Britton S, Cheng Q, Sutherland CJ, McCarthy JS. A simple, high-throughput, colourimetric, field applicable loop-mediated isothermal amplification (HtLAMP) assay for malaria elimination. Malar J. 2015;14:335.CrossRef
20.
go back to reference Lucchi NW, Narayanan J, Karell MA, Xayavong M, Kariuki S, DaSilva AJ, et al. Molecular diagnosis of malaria by photo-induced electron transfer fluorogenic primers: PET-PCR. PLoS ONE. 2013;8:e56677.CrossRef Lucchi NW, Narayanan J, Karell MA, Xayavong M, Kariuki S, DaSilva AJ, et al. Molecular diagnosis of malaria by photo-induced electron transfer fluorogenic primers: PET-PCR. PLoS ONE. 2013;8:e56677.CrossRef
21.
go back to reference Yamamura M, Makimura K, Ota Y. Evaluation of a new rapid molecular diagnostic system for Plasmodium falciparum combined with DNA filter paper, loop-mediated isothermal amplification, and melting curve analysis. Jpn J Infect Dis. 2009;62:20–5.PubMed Yamamura M, Makimura K, Ota Y. Evaluation of a new rapid molecular diagnostic system for Plasmodium falciparum combined with DNA filter paper, loop-mediated isothermal amplification, and melting curve analysis. Jpn J Infect Dis. 2009;62:20–5.PubMed
22.
go back to reference Patel JC, Oberstaller J, Xayavong M, Narayanan J, DeBarry JD, Srinivasamoorthy G, et al. Real-time loop-mediated isothermal amplification (RealAmp) for the species-specific identification of Plasmodium vivax. PLoS ONE. 2013;8:e54986.CrossRef Patel JC, Oberstaller J, Xayavong M, Narayanan J, DeBarry JD, Srinivasamoorthy G, et al. Real-time loop-mediated isothermal amplification (RealAmp) for the species-specific identification of Plasmodium vivax. PLoS ONE. 2013;8:e54986.CrossRef
23.
go back to reference Akerele D, Ljolje D, Talundzic E, Udhayakumar V, Lucchi NW. Molecular diagnosis of Plasmodium ovale by photo-induced electron transfer fluorogenic primers: PET-PCR. PLoS One. 2017;12:e0179178.CrossRef Akerele D, Ljolje D, Talundzic E, Udhayakumar V, Lucchi NW. Molecular diagnosis of Plasmodium ovale by photo-induced electron transfer fluorogenic primers: PET-PCR. PLoS One. 2017;12:e0179178.CrossRef
Metadata
Title
Field evaluation of malaria malachite green loop-mediated isothermal amplification in health posts in Roraima state, Brazil
Authors
Heather M. Kudyba
Jaime Louzada
Dragan Ljolje
Karl A. Kudyba
Vasant Muralidharan
Joseli Oliveira-Ferreira
Naomi W. Lucchi
Publication date
01-12-2019
Publisher
BioMed Central
Published in
Malaria Journal / Issue 1/2019
Electronic ISSN: 1475-2875
DOI
https://doi.org/10.1186/s12936-019-2722-1

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