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

Alexandra Martín-Ramírez Malaria and Emerging Parasitic Diseases Laboratory, National Microbiology Centre, Instituto de Salud Carlos III, Madrid, Spain;

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Marta Lanza-Suárez Malaria and Emerging Parasitic Diseases Laboratory, National Microbiology Centre, Instituto de Salud Carlos III, Madrid, Spain;

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Carlota Muñoz-García Malaria and Emerging Parasitic Diseases Laboratory, National Microbiology Centre, Instituto de Salud Carlos III, Madrid, Spain;

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Shamilah R. Hisam Parasitology Unit, Infectious Disease Research Centre, Institute for Medical Research, National Institute of Health, Ministry of Health Malaysia, Setia Alam, Selangor, Malaysia;

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Ana Perez-Ayala Department of Clinical Microbiology, Hospital Universitario 12 de Octubre, Madrid, Spain

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José M. Rubio Malaria and Emerging Parasitic Diseases Laboratory, National Microbiology Centre, Instituto de Salud Carlos III, 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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