Title
Comparison of real time and malachite-green based loop-mediated isothermal amplification assays for the detection of Plasmodium vivax and P. falciparum
Date Issued
01 June 2020
Access level
open access
Resource Type
journal article
Author(s)
U.S. Naval Medical Research
U.S. Naval Medical Research
Publisher(s)
Public Library of Science
Abstract
The current context of malaria elimination requires urgent development and implementation of highly sensitive and specific methods for prompt detection and treatment of malaria parasites. Such methods should overcome current delays in diagnosis, allow the detection of low-density infections and address the difficulties in accessing remote endemic communities. In this study, we assessed the performance of the RealAmp and malachite-green loop mediated isothermal amplification (MG-LAMP) methodologies, using microscopy and conventional nested-PCR as reference techniques. Both LAMP techniques were performed for Plasmodium genus, P. falciparum, and P. vivax identification using 136 whole blood samples collected from three communities located in the Peruvian Amazon basin. Turnaround time and costs of performing the LAMP assays were estimated and compared to that of microscopy and nested-PCR. Using nested-PCR as reference standard, we calculated the sensitivity, specificity and 95% confidence interval (CI) for all methods. RealAmp had a sensitivity of 92% (95% CI: 85-96.5%) and specificity of 100% (95% CI: 89.1-100%) for species detection; sensitivity and specificity of MG-LAMP were 94% (95% CI: 87.5-97.8%) and 100% (89.1-100%), respectively. Whereas microscopy showed 88.1% sensitivity (95% CI: 80.2-93.7%) and 100% specificity (95%: 89.1-100%). The turnaround time and costs of performing the LAMP assays were lower compared to those associated with nested-PCR but higher than those associated with microscopy. The two LAMP assays were shown to be more sensitive and simple to implement than microscopy. Both LAMP methodologies could be used as large-scale screening tests, but the MG-LAMP assay uses a simple, portable heat-block while the RealAmp requires a RealAmp machine or a real-time PCR machine. This makes the MG-LAMP an appropriate choice for malaria surveillance studies in endemic sites. Use of LAMP tests in active case detection of Plasmodium parasites could help to detect positive malaria cases early.
Volume
15
Issue
6
Language
English
OCDE Knowledge area
Parasitología
Scopus EID
2-s2.0-85086355471
PubMed ID
Source
PLoS ONE
ISSN of the container
19326203
Sponsor(s)
This work was supported by the US DoD Armed Forces Health Surveillance Center and its Global Emerging Infections Surveillance and Response System section (AFHSC/GEIS), PROMIS ID P0106_18_N6_02. We would like to thank Dr. Venkatachalam Udhayakumar for his support in reviewing the article and the laboratory team in Iquitos for their continuous hard work in collecting samples. Disclaimer: The views expressed in this article are those of the authors and do not necessarily reflect the official policy or position of the Department of the Navy, Department of Defense, nor the U.S. Government nor do they represent the official position of the Centers for Disease Control and Prevention. Use of trade names is for identification only and does not imply endorsement by [the Centers for Disease Control and Prevention/the Agency for Toxic Substances and Disease Registry], the Public Health Service, or the U.S. Department of Health and Human Services.
Sources of information: Directorio de Producción Científica Scopus