NINA-LAMP compared to microscopy, RDT, and nested PCR for the detection of imported malaria

Mohon, Abu Naser; Lee, Lydia Da-Yeong; Bayih, Abebe Genetu; Folefoc, Asongna; Guelig, Dylan; Burton, Robert P.; LaBarre, Paul; Chan, Wilson; Meatherall, Bonnie; Pillai, Dylan R.

doi:10.1016/j.diagmicrobio.2015.11.009

Cited by 40 publications

(45 citation statements)

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“…LAMP-based methods for the detection of malaria have been available on a researchuse-only basis for over a decade (28). Results from studies performed both in settings where malaria is endemic and where it is not endemic have suggested that LAMP is more sensitive than rapid diagnostic tests (RDTs) such as the BinaxNOW malaria test, especially for the non-falciparum species (13,17,30,35). LAMP appears to perform on an equal footing with other molecular techniques such as nested PCR (35).…”

Section: Discussionmentioning

confidence: 99%

“…Results from studies performed both in settings where malaria is endemic and where it is not endemic have suggested that LAMP is more sensitive than rapid diagnostic tests (RDTs) such as the BinaxNOW malaria test, especially for the non-falciparum species (13,17,30,35). LAMP appears to perform on an equal footing with other molecular techniques such as nested PCR (35). LAMP may be especially useful for the screening of low-level parasitemia due to its analytical sensitivity (17).…”

Section: Discussionmentioning

confidence: 99%

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Detection of Plasmodium Infection by the illumigene Malaria Assay Compared to Reference Microscopy and Real-Time PCR

et al. 2017

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Malaria is one of the leading causes of infectious disease in travelers returning from the tropics. The diagnosis of malaria is typically performed by examining Giemsa-stained thick and thin peripheral blood smears, which is time consuming, labor intensive, and requires high levels of proficiency. Alternatively, loop-mediated isothermal amplification (LAMP) is a new molecular method, which is rapid, sensitive, and requires less capital equipment and technological training. We conducted a retrospective study comparing two formats of a commercial LAMP assay (Meridian gene malaria [M] and malaria Plus [MP]) versus reference microscopy on archived blood specimens ( = 140) obtained from unique returning travelers suspected of having malaria. Discrepant results were resolved by either repeat testing or a laboratory developed ultrasensitive real-time PCR method. On initial testing, the Meridian gene M and MP kits had sensitivities of 97.3% (95% confidence interval [CI], 90.7 to 99.7%) and 100.0% (95.1 to 100.0%) and specificities of 93.8% (84.8 to 98.3%) and 91.5% (81.3 to 97.2%), respectively, versus reference microscopy. We project a significant cost reduction in low prevalence settings where malaria is not endemic with LAMP-based malaria screening given the excellent negative predictive value achieved with LAMP.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Detection of Plasmodium Infection by the illumigene Malaria Assay Compared to Reference Microscopy and Real-Time PCR

et al. 2017

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“…To further facilitate the field use of isothermal amplification techniques in resource-limited settings, PATH developed an inexpensive, highly efficient, and electricity-free non-instrumented nucleic acid amplification (NINA) system that is based on an exothermic reaction between common salt and Mg-Fe mechanical alloy [50–55]. We describe here the development of a novel RT-LAMP technique that is based on detection of a high-abundant Plasmodium falciparum transcript in parasite RNA extracts or infected whole blood lysates using commercially available single enzyme systems and the NINA approach.…”

Section: Introductionmentioning

confidence: 99%

A Field-Tailored Reverse Transcription Loop-Mediated Isothermal Assay for High Sensitivity Detection of Plasmodium falciparum Infections

Kemleu¹,

Guelig²,

Moukoko³

et al. 2016

PLoS ONE

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Highly sensitive and field deployable molecular diagnostic tools are critically needed for detecting submicroscopic, yet transmissible levels of malaria parasites prevalent in malaria endemic countries worldwide. A reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed and evaluated in comparison with thick blood smear microscopy, an antigen-based rapid diagnostic test (RDT), and an in-house RT-PCR targeting the same RT-LAMP transcript. The optimized assay detected Plasmodium falciparum infections in as little as 0.25ng of total parasite RNA, and exhibited a detection limit of 0.08 parasites/ μL when tested directly on infected whole blood lysates, or ~0.0008 parasites/ μL when using RNA extracts. Assay positivity was observed as early as eight minutes from initiation of the RT-LAMP and in most cases the reaction was complete before twenty minutes. Clinical evaluation of the assay on 132 suspected malaria cases resulted in a positivity rate of 90% for RT-LAMP using extracted RNA, and 85% when using whole blood lysates. The positivity rates were 70% for P. falciparum-specific RDT, 83% for RT-PCR, and 74% for thick blood smear microscopy (Mean parasite density = 36,986 parasites/ μL). Concordance rates between the developed RT-LAMP and comparator tests were greater than 75%, the lowest being with light microscopy (78%, McNemar’s test: P = 0.0002), and the highest was with RT-PCR (87%, McNemar’s test: P = 0.0523). Compared to reference RT-PCR, assay sensitivity was 90% for RT-LAMP on whole blood, and 96% for RT-LAMP using corresponding RNA extracts. Electricity-free heaters were further developed and evaluated in comparison with a battery-operated isothermal amplification machine for use with the developed test in resource-limited settings. Taken together, the data highlight the benefits of targeting high abundant RNA transcripts in molecular diagnosis, as well as the potential usefulness of the developed RT-LAMP-assay in malaria diagnosis in low to high parasite density settings.

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“…Un estudio realizado en Colombia indica que a través de este método de microscopía, la sensibilidad es 93% y especificidad es 98,7% para P. falciparum, mientras que para P. vivax, la sensibilidad es 98% y la especificidad es 97, 9% [29]. Dentro de las dificultades que presenta este método es que solamente detecta parasitemias mayores a 50 parásitos/μL, y no permite detectar coinfecciones; esta incapacidad de detectar infecciones con baja carga de pará-sitos sanguíneos, conduce al no tratamiento de los pacientes y por ende al progreso de la enfermedad [30,31].…”

Section: Métodos Microscópicosunclassified

Malaria, enfermedad tropical de múltiples métodos diagnósticos

Moreno

Sánchez

Giraldo

et al. 2017

archmed

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La malaria es una enfermedad causada por el parasito de género Plasmodium, que se transmite mediante la picadura de la hembra del mosquito Anopheles. Entre los años 2000 a 2015 se han reportado 212 millones de casos a nivel mundial. En Colombia, la malaria es considerada un problema de salud pública, se estima que al menos el 70% del territorio cuenta con las condiciones ecológicas necesarias para la transmisión del parasito. Para evitar el aumento en el número de casos de malaria, se utilizan 3 pilares de prevención: evitar la exposición y realizar control del vector, quimioprofilaxis y diagnóstico oportuno. Después de este último, se debe brindar el tratamiento, cuyos objetivos son: curación clínica del paciente, curación radical de la infección y control de la transmisión. Para el diagnóstico, existen diferentes métodos, los cuales se evalúan y diferencian de acuerdo a la sensibilidad y especificidad; con base en esto, se define el Gold Standard, como prueba de referencia. En la actualidad, el Gold Standard para el diagnóstico de malaria es un método microscópico: gota gruesa. Sin embargo, existen otros métodos: extendido de sangre periférica, métodos moleculares y métodos serológicos.

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NINA-LAMP compared to microscopy, RDT, and nested PCR for the detection of imported malaria

Cited by 40 publications

References 40 publications

Detection of Plasmodium Infection by the illumigene Malaria Assay Compared to Reference Microscopy and Real-Time PCR

Detection of Plasmodium Infection by the illumigene Malaria Assay Compared to Reference Microscopy and Real-Time PCR

A Field-Tailored Reverse Transcription Loop-Mediated Isothermal Assay for High Sensitivity Detection of Plasmodium falciparum Infections

Malaria, enfermedad tropical de múltiples métodos diagnósticos

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