Skip to main content
Top
Published in: Malaria Journal 1/2020

Open Access 01-12-2020 | Malaria | Research

Evaluation of the colorimetric malachite green loop-mediated isothermal amplification (MG-LAMP) assay for the detection of malaria species at two different health facilities in a malaria endemic area of western Kenya

Authors: James Gachugia, Winnie Chebore, Kephas Otieno, Caroline Wangari Ngugi, Adano Godana, Simon Kariuki

Published in: Malaria Journal | Issue 1/2020

Login to get access

Abstract

Background

Prompt diagnosis and effective malaria treatment is a key strategy in malaria control. However, the recommended diagnostic methods, microscopy and rapid diagnostic tests (RDTs), are not supported by robust quality assurance systems in endemic areas. This study compared the performance of routine RDTs and smear microscopy with a simple molecular-based colorimetric loop-mediated isothermal amplification (LAMP) at two different levels of the health care system in a malaria-endemic area of western Kenya.

Methods

Patients presenting with clinical symptoms of malaria at Rota Dispensary (level 2) and Siaya County Referral Hospital (level 4) were enrolled into the study after obtaining written informed consent. Capillary blood was collected to test for malaria by RDT and microscopy at the dispensary and county hospital, and for preparation of blood smears and dried blood spots (DBS) for expert microscopy and real-time polymerase chain reaction (RT-PCR). Results of the routine diagnostic tests were compared with those of malachite green loop-mediated isothermal amplification (MG-LAMP) performed at the two facilities.

Results

A total of 264 participants were enrolled into the study. At the dispensary level, the positivity rate by RDT, expert microscopy, MG-LAMP and RT-PCR was 37%, 30%, 44% and 42%, respectively, and 42%, 43%, 57% and 43% at the county hospital. Using RT-PCR as the reference test, the sensitivity of RDT and MG-LAMP was 78.1% (CI 67.5–86.4) and 82.9% (CI 73.0–90.3) at Rota dispensary. At Siaya hospital the sensitivity of routine microscopy and MG-LAMP was 83.3% (CI 65.3–94.4) and 93.3% (CI 77.9–99.2), respectively. Compared to MG-LAMP, there were 14 false positives and 29 false negatives by RDT at Rota dispensary and 3 false positives and 13 false negatives by routine microscopy at Siaya Hospital.

Conclusion

MG-LAMP is more sensitive than RDTs and microscopy in the detection of malaria parasites at public health facilities and might be a useful quality control tool in resource-limited settings.
Literature
1.
go back to reference WHO. World malaria report 2018. Geneva, World Health Organization, 2018. WHO. World malaria report 2018. Geneva, World Health Organization, 2018.
2.
go back to reference WHO. Global Technical Strategy for Malaria 2016–2030. Geneva, World Health Organization; 2015. WHO. Global Technical Strategy for Malaria 2016–2030. Geneva, World Health Organization; 2015.
3.
go back to reference WHO. Malaria. Geneva, World Health Organization, 2017. WHO. Malaria. Geneva, World Health Organization, 2017.
4.
go back to reference Bastiaens GJ, Bousema T, Leslie T. Scale-up of malaria rapid diagnostic tests and artemisinin-based combination therapy: challenges and perspectives in sub-Saharan Africa. PLoS Med. 2014;11:e1001590. PubMedPubMedCentralCrossRef Bastiaens GJ, Bousema T, Leslie T. Scale-up of malaria rapid diagnostic tests and artemisinin-based combination therapy: challenges and perspectives in sub-Saharan Africa. PLoS Med. 2014;11:e1001590. PubMedPubMedCentralCrossRef
5.
go back to reference Kachur SP. Beyond “test and treat” – malaria diagnosis for improved pediatric fever management in sub-Saharan Africa by Emily White Johansson. Glob Health Action. 2016;9:34416.PubMedCrossRef Kachur SP. Beyond “test and treat” – malaria diagnosis for improved pediatric fever management in sub-Saharan Africa by Emily White Johansson. Glob Health Action. 2016;9:34416.PubMedCrossRef
6.
go back to reference Sori G, Zewdie O, Tadele G, Samuel A. External quality assessment of malaria microscopy diagnosis in selected health facilities in Western Oromia. Ethiopia Malar J. 2018;17:233.PubMedCrossRef Sori G, Zewdie O, Tadele G, Samuel A. External quality assessment of malaria microscopy diagnosis in selected health facilities in Western Oromia. Ethiopia Malar J. 2018;17:233.PubMedCrossRef
7.
go back to reference Wanja E, Achilla RA, Obare P, Adeny R, Moseti C, Otieno V, et al. Evaluation of a laboratory quality assurance pilot programme for malaria diagnostics in low-transmission areas of Kenya, 2013. Malar J. 2017;16:221.PubMedPubMedCentralCrossRef Wanja E, Achilla RA, Obare P, Adeny R, Moseti C, Otieno V, et al. Evaluation of a laboratory quality assurance pilot programme for malaria diagnostics in low-transmission areas of Kenya, 2013. Malar J. 2017;16:221.PubMedPubMedCentralCrossRef
8.
go back to reference Odhiambo F, Buff AM, Moranga C, Moseti CM, Wesongah JO, Lowther SA, et al. Factors associated with malaria microscopy diagnostic performance following a pilot quality-assurance programme in health facilities in malaria low-transmission areas of Kenya, 2014. Malar J. 2017;16:371.PubMedPubMedCentralCrossRef Odhiambo F, Buff AM, Moranga C, Moseti CM, Wesongah JO, Lowther SA, et al. Factors associated with malaria microscopy diagnostic performance following a pilot quality-assurance programme in health facilities in malaria low-transmission areas of Kenya, 2014. Malar J. 2017;16:371.PubMedPubMedCentralCrossRef
9.
go back to reference Jemere KA, Melaku MY, Jemeber TH, Abate MA. Performance evaluation of laboratory professionals on malaria microscopy at health facilities in Bahir Dar city administration, Northwest Ethiopia. PLoS One. 2018;13. Jemere KA, Melaku MY, Jemeber TH, Abate MA. Performance evaluation of laboratory professionals on malaria microscopy at health facilities in Bahir Dar city administration, Northwest Ethiopia. PLoS One. 2018;13.
10.
go back to reference Beshir KB, Sepúlveda N, Bharmal J, Robinson A, Mwanguzi J, Busula AO, et al. Plasmodium falciparum parasites with histidine-rich protein 2 (pfhrp2) and pfhrp3 gene deletions in two endemic regions of Kenya. Sci Rep. 2017;7:14718.PubMedPubMedCentralCrossRef Beshir KB, Sepúlveda N, Bharmal J, Robinson A, Mwanguzi J, Busula AO, et al. Plasmodium falciparum parasites with histidine-rich protein 2 (pfhrp2) and pfhrp3 gene deletions in two endemic regions of Kenya. Sci Rep. 2017;7:14718.PubMedPubMedCentralCrossRef
11.
go back to reference WHO. Malaria rapid diagnostic test performance. Results of WHO product testing of malaria RDTs: Round 3 (2010–2011). Geneva, World Health Organization; 2011. WHO. Malaria rapid diagnostic test performance. Results of WHO product testing of malaria RDTs: Round 3 (2010–2011). Geneva, World Health Organization; 2011.
12.
go back to reference WHO. Global Malaria Programme: Policy brief on malaria diagnostics in low-transmission settings. Geneva: World Health Organization; 2014. WHO. Global Malaria Programme: Policy brief on malaria diagnostics in low-transmission settings. Geneva: World Health Organization; 2014.
13.
go back to reference Berzosa P, de Lucio A, Romay‑Barja M, Herrador Z, González V, García. L, et al. Comparison of three diagnostic methods (microscopy, RDT, and PCR) for the detection of malaria parasites in representative samples from Equatorial Guinea. Malar J. 2018;17:333. Berzosa P, de Lucio A, Romay‑Barja M, Herrador Z, González V, García. L, et al. Comparison of three diagnostic methods (microscopy, RDT, and PCR) for the detection of malaria parasites in representative samples from Equatorial Guinea. Malar J. 2018;17:333.
14.
go back to reference Mahmud Y-LL, Meng-Yee L, Mun-Yik F, Jenarun J, Rohela. Loop-mediated isothermal amplification assay for identification of five human Plasmodium species in Malaysia. Am J Trop Med Hyg. 2016;94:336–9. Mahmud Y-LL, Meng-Yee L, Mun-Yik F, Jenarun J, Rohela. Loop-mediated isothermal amplification assay for identification of five human Plasmodium species in Malaysia. Am J Trop Med Hyg. 2016;94:336–9.
15.
go back to reference Ministry of Health. National guidelines for the diagnosis, treatment and prevention of malaria in Kenya. Nairobi, 2016. Ministry of Health. National guidelines for the diagnosis, treatment and prevention of malaria in Kenya. Nairobi, 2016.
16.
go back to reference Kapesa A, Kweka EJ, Zhou G, Atieli HE, Kamugisha E, Mazigo HD, et al. Utility of passive malaria surveillance in hospitals as a surrogate to community infection transmission dynamics in western Kenya. Arch Public Health. 2018;76:39.PubMedPubMedCentralCrossRef Kapesa A, Kweka EJ, Zhou G, Atieli HE, Kamugisha E, Mazigo HD, et al. Utility of passive malaria surveillance in hospitals as a surrogate to community infection transmission dynamics in western Kenya. Arch Public Health. 2018;76:39.PubMedPubMedCentralCrossRef
17.
go back to reference Bayoh MN, Walker ED, Kosgei J, Ombok M, Olang GB, Githeko AK, et al. Persistently high estimates of late night, indoor exposure to malaria vectors despite high coverage of insecticide treated nets. Parasit Vectors. 2014;7:380.PubMedCrossRef Bayoh MN, Walker ED, Kosgei J, Ombok M, Olang GB, Githeko AK, et al. Persistently high estimates of late night, indoor exposure to malaria vectors despite high coverage of insecticide treated nets. Parasit Vectors. 2014;7:380.PubMedCrossRef
18.
go back to reference WHO. Basic Malaria Microscopy: Part 1. Learner's Guide. 2nd edition. Geneva, World Health Organization; 2010. WHO. Basic Malaria Microscopy: Part 1. Learner's Guide. 2nd edition. Geneva, World Health Organization; 2010.
19.
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. 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.
20.
go back to reference McConnico DB, Alessandra EF, Michael CH, John DB, Caitlin C. Development of new malaria diagnostics: matching performance and need. Malar J. 2016;15:406.PubMedPubMedCentralCrossRef McConnico DB, Alessandra EF, Michael CH, John DB, Caitlin C. Development of new malaria diagnostics: matching performance and need. Malar J. 2016;15:406.PubMedPubMedCentralCrossRef
21.
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.PubMedPubMedCentralCrossRef 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.PubMedPubMedCentralCrossRef
22.
go back to reference WHO-FIND-CDC. Methods manual for product testing of malaria rapid diagnostic tests. Geneva, World Health Organization; 2018. WHO-FIND-CDC. Methods manual for product testing of malaria rapid diagnostic tests. Geneva, World Health Organization; 2018.
23.
go back to reference Watson OJ, Sumner KM, Janko M, Goel V, Winskill P, Slater HC, et al. False-negative malaria rapid diagnostic test results and their impact on community-based malaria surveys in sub-Saharan Africa. BMJ Glob Health. 2019;4:e001582.PubMedPubMedCentralCrossRef Watson OJ, Sumner KM, Janko M, Goel V, Winskill P, Slater HC, et al. False-negative malaria rapid diagnostic test results and their impact on community-based malaria surveys in sub-Saharan Africa. BMJ Glob Health. 2019;4:e001582.PubMedPubMedCentralCrossRef
24.
go back to reference Shaio P-WL, Dar-Der J, Chia-Tai L, Herodes SR, Virgilio EdR, Lin IF, et al. Application of loop-mediated isothermal amplification for malaria diagnosis during a follow-up study in São Tomé. Malar J. 2012;11:408. Shaio P-WL, Dar-Der J, Chia-Tai L, Herodes SR, Virgilio EdR, Lin IF, et al. Application of loop-mediated isothermal amplification for malaria diagnosis during a follow-up study in São Tomé. Malar J. 2012;11:408.
25.
go back to reference Bwire GM, Ngasala B, Kilonzi M, Mikomangwa WP, Fatuma F. Felician FF, et al. Diagnostic performance of CareStart™ malaria HRP2/pLDH test in comparison with standard microscopy for detection of uncomplicated malaria infection among symptomatic patients, Eastern Coast of Tanzania. Malar J. 2019;18:354. Bwire GM, Ngasala B, Kilonzi M, Mikomangwa WP, Fatuma F. Felician FF, et al. Diagnostic performance of CareStart™ malaria HRP2/pLDH test in comparison with standard microscopy for detection of uncomplicated malaria infection among symptomatic patients, Eastern Coast of Tanzania. Malar J. 2019;18:354.
26.
go back to reference Mfuh KO, Achonduh-Atijegbe OA, Bekindaka ON, Esemu LF, Mbakop CD, Gandhi K, et al. A comparison of thick-film microscopy, rapid diagnostic test, and polymerase chain reaction for accurate diagnosis of Plasmodium falciparum malaria. Malar J. 2019;18:73.PubMedPubMedCentralCrossRef Mfuh KO, Achonduh-Atijegbe OA, Bekindaka ON, Esemu LF, Mbakop CD, Gandhi K, et al. A comparison of thick-film microscopy, rapid diagnostic test, and polymerase chain reaction for accurate diagnosis of Plasmodium falciparum malaria. Malar J. 2019;18:73.PubMedPubMedCentralCrossRef
27.
go back to reference Waitumbi JN, Gerlach J, Afonina I, Anyona SB, Koros JN, Siangla J, et al. Malaria prevalence defined by microscopy, antigen detection, DNA amplification and total nucleic acid amplification in a malaria-endemic region during the peak malaria transmission season. Trop Med Int Health. 2011;16:786–93.PubMedCrossRef Waitumbi JN, Gerlach J, Afonina I, Anyona SB, Koros JN, Siangla J, et al. Malaria prevalence defined by microscopy, antigen detection, DNA amplification and total nucleic acid amplification in a malaria-endemic region during the peak malaria transmission season. Trop Med Int Health. 2011;16:786–93.PubMedCrossRef
28.
go back to reference Masanja IM, McMorrow ML, Maganga MB, Sumari D, Udhayakumar V, McElroy PD, et al. Quality assurance of malaria rapid diagnostic tests used for routine patient care in rural Tanzania: microscopy versus real-time polymerase chain reaction. Malar J. 2015;14:85.PubMedPubMedCentralCrossRef Masanja IM, McMorrow ML, Maganga MB, Sumari D, Udhayakumar V, McElroy PD, et al. Quality assurance of malaria rapid diagnostic tests used for routine patient care in rural Tanzania: microscopy versus real-time polymerase chain reaction. Malar J. 2015;14:85.PubMedPubMedCentralCrossRef
29.
go back to reference Wanja EK, Moranga N, Hickman C, Johnson M, Moseti J, Anova C, et al. Field evaluation of diagnostic performance of malaria rapid diagnostic tests in western Kenya. Malar J. 2016;15:456.PubMedPubMedCentralCrossRef Wanja EK, Moranga N, Hickman C, Johnson M, Moseti J, Anova C, et al. Field evaluation of diagnostic performance of malaria rapid diagnostic tests in western Kenya. Malar J. 2016;15:456.PubMedPubMedCentralCrossRef
30.
go back to reference Tambo M, Mwinga M, Mumbengegwi DR. Loop-mediated isothermal amplification (LAMP) and Polymerase Chain Reaction (PCR) as quality assurance tools for Rapid Diagnostic Test (RDT) malaria diagnosis in Northern Namibia. PLoS ONE. 2018;13:e0206848.PubMedPubMedCentralCrossRef Tambo M, Mwinga M, Mumbengegwi DR. Loop-mediated isothermal amplification (LAMP) and Polymerase Chain Reaction (PCR) as quality assurance tools for Rapid Diagnostic Test (RDT) malaria diagnosis in Northern Namibia. PLoS ONE. 2018;13:e0206848.PubMedPubMedCentralCrossRef
31.
go back to reference ACTwatch Group, Musuva A, Ejersa W, Kiptui R, Memusi D, Abwao E. The malaria testing and treatment landscape in Kenya: results from a nationally representative survey among the public and private sector in 2016. Malar J. 2017;16:494. ACTwatch Group, Musuva A, Ejersa W, Kiptui R, Memusi D, Abwao E. The malaria testing and treatment landscape in Kenya: results from a nationally representative survey among the public and private sector in 2016. Malar J. 2017;16:494.
32.
go back to reference Hailu HA, Shiferaw MB, Demeke L, Derebe MM, Gelaw ZD, Emiru MA, et al. External quality assessment of malaria microscopy diagnosis among public health facilities in West Amhara Region. Ethiopia BMC Res Notes. 2017;10:764.PubMedCrossRef Hailu HA, Shiferaw MB, Demeke L, Derebe MM, Gelaw ZD, Emiru MA, et al. External quality assessment of malaria microscopy diagnosis among public health facilities in West Amhara Region. Ethiopia BMC Res Notes. 2017;10:764.PubMedCrossRef
33.
go back to reference Beshir KB, Sutherland CJ, Sawa P, Drakeley CJ, Okell L, Mweresa CK, et al. Residual Plasmodium falciparum parasitemia in Kenyan children after artemisinin-combination therapy is associated with increased transmission to mosquitoes and parasite recurrence. J Infect Dis. 2013;208:2017–24.PubMedPubMedCentralCrossRef Beshir KB, Sutherland CJ, Sawa P, Drakeley CJ, Okell L, Mweresa CK, et al. Residual Plasmodium falciparum parasitemia in Kenyan children after artemisinin-combination therapy is associated with increased transmission to mosquitoes and parasite recurrence. J Infect Dis. 2013;208:2017–24.PubMedPubMedCentralCrossRef
34.
go back to reference WHO. Guidelines for the treatment of malaria. 3rd Edn. Geneva, World Health Organization; 2015. WHO. Guidelines for the treatment of malaria. 3rd Edn. Geneva, World Health Organization; 2015.
35.
go back to reference Oriero EC, Okebe J, Jacobs J, Van geertruyden J-P, Nwakanma D, D’Alessandro U. Diagnostic performance of a novel loop-mediated isothermal amplification (LAMP) assay targeting the apicoplast genome for malaria diagnosis in a field setting in sub-Saharan Africa. Malar J. 2015;14:396. Oriero EC, Okebe J, Jacobs J, Van geertruyden J-P, Nwakanma D, D’Alessandro U. Diagnostic performance of a novel loop-mediated isothermal amplification (LAMP) assay targeting the apicoplast genome for malaria diagnosis in a field setting in sub-Saharan Africa. Malar J. 2015;14:396.
36.
go back to reference Morris U, Khamis M, Aydin-Schmidt B, Abass AK, Msellem MI, Nassor MH, et al. Field deployment of loop-mediated isothermal amplification for centralized mass-screening of asymptomatic malaria in Zanzibar: A pre-elimination setting. Malar J. 2015;14:205.PubMedPubMedCentralCrossRef Morris U, Khamis M, Aydin-Schmidt B, Abass AK, Msellem MI, Nassor MH, et al. Field deployment of loop-mediated isothermal amplification for centralized mass-screening of asymptomatic malaria in Zanzibar: A pre-elimination setting. Malar J. 2015;14:205.PubMedPubMedCentralCrossRef
37.
go back to reference Kudyba HM, Louzada J, Ljolje D, Kudyba KA, Muralidharan V, Oliveira-Ferreira J, et al. Field evaluation of malaria malachite green loop-mediated isothermal amplification in health posts in Roraima state. Brazil Malar J. 2019;18:98.PubMedCrossRef Kudyba HM, Louzada J, Ljolje D, Kudyba KA, Muralidharan V, Oliveira-Ferreira J, et al. Field evaluation of malaria malachite green loop-mediated isothermal amplification in health posts in Roraima state. Brazil Malar J. 2019;18:98.PubMedCrossRef
38.
go back to reference Cheaveau J, Nguyen H, Chow B, Marasinghe D, Mohon AN, Yuan H, et al. Clinical validation of a commercial LAMP test for ruling out malaria in returning travelers: A prospective diagnostic trial. Open Forum Infect Dis. 2018;5:11. Cheaveau J, Nguyen H, Chow B, Marasinghe D, Mohon AN, Yuan H, et al. Clinical validation of a commercial LAMP test for ruling out malaria in returning travelers: A prospective diagnostic trial. Open Forum Infect Dis. 2018;5:11.
39.
go back to reference Vincent JP, Komaki-Yasuda K, Iwagami M, Kawai S, Kano S. Combination of PURE-DNA extraction and LAMP-DNA amplification methods for accurate malaria diagnosis on dried blood spots. Malar J. 2018;17:373.PubMedPubMedCentralCrossRef Vincent JP, Komaki-Yasuda K, Iwagami M, Kawai S, Kano S. Combination of PURE-DNA extraction and LAMP-DNA amplification methods for accurate malaria diagnosis on dried blood spots. Malar J. 2018;17:373.PubMedPubMedCentralCrossRef
40.
go back to reference Björkman JC, Berit A-S, Iveth JG, David B, Elin E, Majda HN, 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.PubMedPubMedCentralCrossRef Björkman JC, Berit A-S, Iveth JG, David B, Elin E, Majda HN, 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.PubMedPubMedCentralCrossRef
41.
go back to reference Kabaghe AN, Phiri MD, Phiri KS, van Vugt M. Challenges in implementing uncomplicated malaria treatment in children: a health facility survey in rural Malawi. Malar J. 2017;16:419.PubMedPubMedCentralCrossRef Kabaghe AN, Phiri MD, Phiri KS, van Vugt M. Challenges in implementing uncomplicated malaria treatment in children: a health facility survey in rural Malawi. Malar J. 2017;16:419.PubMedPubMedCentralCrossRef
42.
go back to reference Diallo MA, Diongue K, Seck MC, Ndiaye M, Diallo I, Diedhiou Y, et al. Quality control of malaria microscopy reveals misdiagnosed non-falciparum species and other microscopically detectable pathogens in Senegal. Ann Clin Microbiol Antimicrob. 2018;17:8.PubMedPubMedCentralCrossRef Diallo MA, Diongue K, Seck MC, Ndiaye M, Diallo I, Diedhiou Y, et al. Quality control of malaria microscopy reveals misdiagnosed non-falciparum species and other microscopically detectable pathogens in Senegal. Ann Clin Microbiol Antimicrob. 2018;17:8.PubMedPubMedCentralCrossRef
43.
go back to reference Zimmerman PA, Howes RE. Malaria diagnosis for malaria elimination. Curr Opin Infect Dis. 2015;28:446–54.PubMedCrossRef Zimmerman PA, Howes RE. Malaria diagnosis for malaria elimination. Curr Opin Infect Dis. 2015;28:446–54.PubMedCrossRef
Metadata
Title
Evaluation of the colorimetric malachite green loop-mediated isothermal amplification (MG-LAMP) assay for the detection of malaria species at two different health facilities in a malaria endemic area of western Kenya
Authors
James Gachugia
Winnie Chebore
Kephas Otieno
Caroline Wangari Ngugi
Adano Godana
Simon Kariuki
Publication date
01-12-2020
Publisher
BioMed Central
Keywords
Malaria
Plasmodia
Published in
Malaria Journal / Issue 1/2020
Electronic ISSN: 1475-2875
DOI
https://doi.org/10.1186/s12936-020-03397-0

Other articles of this Issue 1/2020

Malaria Journal 1/2020 Go to the issue