• 1.

    WHO , 2020. World Malaria Report 2020—20 Years of Global Progress and Challenges. Geneva, Switzerland: World Health Organization.

  • 2.

    Askling HH, Nilsson J, Tegnell A, Janzon R, Ekdahl K , 2005. Malaria risk in travelers. Emerg Infect Dis 11: 436441.

  • 3.

    European Centre for Disease Prevention and Control , 2021. Malaria. Annual Epidemiological Report for 2019.

  • 4.

    Norman FF et al.2017. Imported malaria in Spain (2009–2016): results from the +REDIVI Collaborative Network. Malar J 16: 407.

  • 5.

    Pérez-Molina JA, López-Polín A, Treviño B, Molina I, Goikoetxea J, Díaz-Menéndez M, Torrús D, Calabuig E, Benito A, López-Vélez R, +Redivi Study Group , 2017. 6-year review of +Redivi: a prospective registry of imported infectious diseases in Spain. J Travel Med 24: 17.

    • Search Google Scholar
    • Export Citation
  • 6.

    Herrador Z, Fernández-Martinez B, Quesada-Cubo V, Diaz-Garcia O, Cano R, Benito A, Gómez-Barroso D , 2019. Imported cases of malaria in Spain: observational study using nationally reported statistics and surveillance data, 2002–2015. Malar J 18: 230.

    • Search Google Scholar
    • Export Citation
  • 7.

    Velasco E, Gomez-Barroso D, Varela C, Diaz O, Cano R , 2017. Non-imported malaria in non-endemic countries: a review of cases in Spain. Malar J 16: 260.

    • Search Google Scholar
    • Export Citation
  • 8.

    Chiodini PL , 2014. Malaria diagnostics: now and the future. Parasitology 141: 18731879.

  • 9.

    Han E-T , 2013. Loop-mediated isothermal amplification test for the molecular diagnosis of malaria. Expert Rev Mol Diagn 13: 205218.

    • Search Google Scholar
    • Export Citation
  • 10.

    Frickmann H, Hinz R, Rojak S, Bonow I, Ruben S, Wegner C, Zielke I, Hagen RM, Tannich E , 2018. Evaluation of automated loop-mediated amplification (LAMP) for routine malaria detection in blood samples of German travelers—a cross-sectional study. Travel Med Infect Dis 24: 2530.

    • Search Google Scholar
    • Export Citation
  • 11.

    Notomi T, Okayami H, Masubuchi H, Yonekawa T, Watanabe K, Amino N, Hase T , 2000. Loop-mediated isothermal amplification of DNA. Nucleic Acids Res 28: e63.

    • Search Google Scholar
    • Export Citation
  • 12.

    Britton S et al.2016. Sensitive detection of Plasmodium vivax using a high-throughput, colourimetric loop mediated isothermal amplification (HtLAMP) platform: a potential novel tool for malaria elimination. PLoS Negl Trop Dis 10: e0004443.

    • Search Google Scholar
    • Export Citation
  • 13.

    Tanner NA, Zhang Y, Evans TC , 2015. Visual detection of isothermal nucleic acid amplification using pH-sensitive dyes. Biotechniques 58: 5968.

    • Search Google Scholar
    • Export Citation
  • 14.

    Nolasco O, Montoya J, Rosales Rosas AL, Barrientos S, Rosanas-Urgell A, Gamboa D , 2021. Multicopy targets for Plasmodium vivax and Plasmodium falciparum detection by colorimetric LAMP. Malar J 20: 225.

    • Search Google Scholar
    • Export Citation
  • 15.

    Lucchi NW, Ljolje D, Silva-Flannery L, Udhayakumar V , 2016. Use of malachite green-loop mediated isothermal amplification for detection of Plasmodium spp. parasites. PLoS One 11: e0151437.

    • Search Google Scholar
    • Export Citation
  • 16.

    Kudyba HM, Louzada J, Ljolje D, Kudyba KA, Muralidharan V, Oliveira-Ferreira J, Lucchi NW , 2019. Field evaluation of malaria malachite green loop-mediated isothermal amplification in health posts in Roraima state, Brazil. Malar J 18: 98.

    • Search Google Scholar
    • Export Citation
  • 17.

    Cook J et al.2015. Loop-mediated isothermal amplification (LAMP) for point-of-care detection of asymptomatic low-density malaria parasite carriers in Zanzibar. Malar J 14: 43.

    • Search Google Scholar
    • Export Citation
  • 18.

    Hopkins H et al.2013. 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 208: 645652.

    • Search Google Scholar
    • Export Citation
  • 19.

    Cuadros J, Pérez-Tanoira R, Prieto-Pérez L, Martin-Martin I, Berzosa P, González V, Tisiano G, Balcha S, Ramos JM, Górgolas M , 2015. Field evaluation of malaria microscopy, rapid malaria tests and loop-mediated isothermal amplification in a rural hospital in south western Ethiopia. PLoS One 10: e0142842.

    • Search Google Scholar
    • Export Citation
  • 20.

    Gachugia J, Chebore W, Otieno K, Ngugi CW, Godana A, Kariuki S , 2020. 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. Malar J 19: 329.

    • Search Google Scholar
    • Export Citation
  • 21.

    Morris U, Aydin-Schmidt B , 2021. Performance and application of commercially available loop-mediated isothermal amplification (LAMP) kits in malaria endemic and non-endemic settings. Diagnostics (Basel) 11: 336.

    • Search Google Scholar
    • Export Citation
  • 22.

    Ljolje D, Abdallah R, Lucchi NW , 2021. Detection of malaria parasites in samples from returning US travelers using the Alethia® Malaria Plus LAMP assay. BMC Res Notes 14: 128.

    • Search Google Scholar
    • Export Citation
  • 23.

    Rubio JM, Post RJ, Docters van Leeuwen WM, Henry MC, Lindergard G, Hommel M , 2002. Alternative polymerase chain reaction method to identify Plasmodium species in human blood samples: the semi-nested multiplex malaria PCR (SnM-PCR). Trans R Soc Trop Med Hyg 96: 199204.

    • Search Google Scholar
    • Export Citation
  • 24.

    Leeflang MMG, Allerberger F , 2019. How to: evaluate a diagnostic test. Clin Microbiol Infect 25: 5459.

  • 25.

    Miguel-Oteo M, Jiram AI, Ta-Tang TH, Lanza M, Hisam S, Rubio JM , 2017. Nested multiplex PCR for identification and detection of human Plasmodium species including Plasmodium knowlesi. Asian Pac J Trop Med 10: 299304.

    • Search Google Scholar
    • Export Citation
  • 26.

    Han E-T, Watanabe R, Sattabongkot J, Khuntirat B, Sirichaisinthop J, Iriko H, Jin L, Takeo S, Tsuboi T , 2007. Detection of four Plasmodium species by genus- and species-specific loop-mediated isothermal amplification for clinical diagnosis. J Clin Microbiol 45: 25212528.

    • Search Google Scholar
    • Export Citation
  • 27.

    Lau Y-L, Lai M-Y, Fong M-Y, Jelip J, Mahmud R , 2016. Loop-mediated isothermal amplification assay for identification of five human Plasmodium species in Malaysia. Am J Trop Med Hyg 94: 336339.

    • Search Google Scholar
    • Export Citation
  • 28.

    Saah AJ , 1997. “Sensitivity” and “specificity” reconsidered: the meaning of these terms in analytical and diagnostic settings. Ann Intern Med 126: 91.

    • Search Google Scholar
    • Export Citation
  • 29.

    Pérez MI, Hervada Vidal X, Naveira-Barbeito G, Silva LC, Fariñas H, Vázquez E, Bacallao J, Mujica O , 2010. EPIDAT programme: uses and perspectives. Rev Panam Salud Publica 27: 8082.

    • Search Google Scholar
    • Export Citation
  • 30.

    Motulsky HJ , 2016. GraphPad Statistics Guide. Available at: https://www.graphpad.com/guides/prism/7/statistics/index/htm. Accessed October 3, 2021.

  • 31.

    Kollenda H, Hagen RM, Hanke M, Rojak S, Hinz R, Wassill L, Poppert S, Tannich E, Frickmann H , 2018. Poor diagnostic performance of a species-specific loop-mediated isothermal amplification (LAMP) platform for malaria. Eur J Microbiol Immunol (Bp) 8: 112118.

    • Search Google Scholar
    • Export Citation
  • 32.

    Imai K et al.2017. A novel diagnostic method for malaria using loop-mediated isothermal amplification (LAMP) and MinIONTM nanopore sequencer. BMC Infect Dis 17: 621.

    • Search Google Scholar
    • Export Citation
  • 33.

    Lucchi NW, Ndiaye D, Britton S, Udhayakumar V , 2018. Expanding the malaria molecular diagnostic options: opportunities and challenges for loop-mediated isothermal amplification tests for malaria control and elimination. Expert Rev Mol Diagn 18: 195203.

    • Search Google Scholar
    • Export Citation
  • 34.

    Nzelu CO, Gomez EA, Cáceres AG, Sakurai T, Martini-Robles L, Uezato H, Mimori T, Katakura K, Hashiguchi Y, Kato H , 2014. Development of a loop-mediated isothermal amplification method for rapid mass-screening of sand flies for Leishmania infection. Acta Trop 132: 16.

    • Search Google Scholar
    • Export Citation
  • 35.

    Ta TH, Hisam S, Lanza M, Jiram AI, Ismail N, Rubio JM , 2014. First case of a naturally acquired human infection with Plasmodium cynomolgi. Malar J 13: 68.

    • Search Google Scholar
    • Export Citation
  • 36.

    Bialasiewicz S, Whiley DM, Nissen MD, Sloots TP , 2007. Impact of competitive inhibition and sequence variation upon the sensitivity of malaria PCR. J Clin Microbiol 45: 16211623.

    • Search Google Scholar
    • Export Citation
  • 37.

    Barazorda KA, Salas CJ, Bishop DK, Lucchi N, Valdivia HO , 2020. Comparison of real time and malachite-green based loop-mediated isothermal amplification assays for the detection of Plasmodium vivax and P. falciparum. PLoS One 15: e0234263.

    • Search Google Scholar
    • Export Citation
  • 38.

    Rubio JM, Benito A, Berzosa PJ, Roche J, Puente S, Subirats M, Lopez-Velez R, Garcia L, Alvar J , 1999. Usefulness of seminested multiplex PCR in surveillance of imported malaria in Spain. J Clin Microbiol 37: 32603264.

    • Search Google Scholar
    • Export Citation
  • 39.

    Landis JR, Koch GG , 1977. An application of hierarchical kappa-type statistics in the assessment of majority agreement among multiple observers. Biometrics 33: 363374.

    • Search Google Scholar
    • Export Citation
  • 40.

    Cuadros J, Martin Ramírez A, González IJ, Ding XC, Perez Tanoira R, Rojo-Marcos G, Gómez-Herruz P, Rubio JM , 2017. LAMP kit for diagnosis of non-falciparum malaria in Plasmodium ovale infected patients. Malar J 16: 20.

    • Search Google Scholar
    • Export Citation
  • 41.

    Centre National de Référence du Paludisme , 2017. Rapport Annuel d’Activité, 2017. Année d’Exercise 2016. Paris, France: CNR du Paludisme.

    • Search Google Scholar
    • Export Citation
  • 42.

    Fradejas I et al.2019. Prevalence of submicroscopic malaria infection in immigrants living in Spain. Malar J 18: 242.

  • 43.

    Lucchi NW et al.2016. Evaluation of the Illumigene Malaria LAMP: a robust molecular diagnostic tool for malaria parasites. Sci Rep 6: 36808.

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Usefulness of Malachite-Green LAMP for Diagnosis of Plasmodium and Five Human Malaria Species in a Nonendemic Setting

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  • 1 Malaria and Emerging Parasitic Diseases Laboratory, National Microbiology Centre, Instituto de Salud Carlos III, Madrid, Spain;
  • | 2 Parasitology Unit, Infectious Disease Research Centre, Institute for Medical Research, National Institute of Health, Ministry of Health Malaysia, Setia Alam, Selangor, Malaysia;
  • | 3 Department of Clinical Microbiology, Hospital Universitario 12 de Octubre, Madrid, Spain
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ABSTRACT.

Molecular methods are necessary to detect low-density malaria infections. The purpose of this study was to assess the diagnostic performance of six malachite-green loop-mediated amplification method (MG-LAMP) assays (MG-LAMP-Pf, MG-LAMP-Pv, MG-LAMP-Po, MG-LAMP-Pm, MG-LAMP-Pk, and MG-LAMP-Pspp) for the species-specific detection of each human Plasmodium, including P. knowlesi, and the Plasmodium genus compared with the nested-multiplex malaria polymerase chain reaction (NM-PCR), using 161 malaria-positive and 274 malaria-negative samples. MG-LAMP-Pspp assay detected the five human Plasmodium species and each species-specific MG-LAMP assay detected only its corresponding species. Sensitivity, specificity, and predictive values of MG-LAMP assays, compared with NM-PCR, were > 90%, except in the case of the MG-LAMP-Pm assay, which dropped to 47%. Limit of detection for MG-LAMP-Pspp assay ranged from 0.1 parasites/µL for P. falciparum to 16.9 parasites/µL for P. malariae samples, and it was similar for the rest of MG-LAMP assays except for the MG-LAMP-Pm assay. Turnaround time was estimated to be 2 hours and 35 minutes for one MG-LAMP assay and 4 hours and 15 minutes if all species-specific MG-LAMP is set up, whereas for the NM-PCR, turnaround time was ∼6 hours and 15 minutes. Costs per determination ranged from 1 to 6 euros for MG-LAMP assays and 5 euros for NM-PCR. Therefore, MG-LAMP assays appear to have good concordance compared with the reference method, except for the MG-LAMP-Pm assay. They can detect low parasitemia and identify malaria species, with lower costs and shorter time to obtain results, and they are suitable tools to be used in endemic and non-endemic countries for malaria detection.

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Author Notes

Address correspondence to José M. Rubio, Malaria & Parasitic Emerging Diseases Laboratory, National Microbiology Center, Instituto de Salud Carlos III, Crta. de Majadahonda a Pozuelo Km. 2. Majadahonda, 28220 Madrid, Spain. E-mail: jmrubio@isciii.es

Financial support: This work was partially supported by the Spanish Strategic Health Action (AESI-ISCIII) Grant Number PI17CIII/00035. The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Alexandra Martín Ramírez is supported by an ISCIII Río Hortega grant.

Authors’ addresses: Alexandra Martín-Ramírez, Marta Lanza-Suárez, Carlota Muñoz-García, and José M. Rubio, Malaria & Parasitic Emerging Diseases Laboratory, National Microbiology Center, Instituto de Salud Carlos III, Madrid, Spain, E-mails: a.martin@isciii.es, mlanza@isciii.es, carlota.mg114@gmail.com, and jmrubio@isciii.es. Shamilah R. Hisam, Parasitology Unit, Infectious Disease Research Centre, Institute for Medical Research, National Institute of Health, Setia Alam, Selangor, Malaysia, E-mail: shamilah@imr.gov.my. Ana Perez-Ayala, Department of Clinical Microbiology, Hospital Universitario 12 de Octubre, Madrid, Spain, E-mail: anpayala@hotmail.com.

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