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Publication Details
AFRICAN RESEARCH NEXUS
SHINING A SPOTLIGHT ON AFRICAN RESEARCH
medicine
Loop-mediated isothermal amplification for rapid and semiquantitative detection of loa loa infection
Journal of Clinical Microbiology, Volume 52, No. 6, Year 2014
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Description
Rapid and accurate tests are currently needed to identify individuals with high levels of Loa loa microfilaria (mf), so that these individuals may be excluded from mass ivermectin administration campaigns against onchocerciasis and lymphatic filariasis being conducted in areas where Onchocerca volvulus, Wuchereria bancrofti, and L. loa are coendemic. To address this need, colorimetric loop-mediated isothermal amplification (LAMP) assays targeting the L. loa-specific gene sequences LLMF72 and LLMF342 were developed for the detection and quantification of L. loa microfilaremia. Both LAMP assays were highly specific (100%) for L. loa infection compared to the absence of infection or infection with related filarial pathogens. The LLMF72-based LAMP assay showed greater analytic sensitivity (limit of detection, 0.1 pg/ml of genomic DNA [gDNA] and/or 5 mf/ml) than the LLMF342-based LAMP assay (10 pg/ml of gDNA and/or 50 mf/ml), and its analytic sensitivity was similar to that of LLMF72-based quantitative PCR (qPCR). A high level of correlation was observed between microfilaria counts as determined by LLMF72-based qPCR and time to positivity by the LAMP assay, and performance measures of sensitivity, specificity, and positive and negative predictive values were similar for both assays when applied to field-collected clinical samples. By simply varying the run time, the LAMP assay was able to accurately distinguish individuals at risk for serious adverse events (SAEs) after exposure to ivermectin, using thresholds of>5,000 mf/ml and>30,000 mf/ml as indicators of increasing levels of risk. In summary, LLMF72 LAMP represents a new molecular diagnostic tool that is readily applicable as a point-of-care method for L. loa microfilarial detection and quantification in resource-limited countries where L. loa infection is endemic. © 2014 American Society for Microbiology. All Rights Reserved.
Authors & Co-Authors
Drame, Papa Makhtar
United States, Bethesda
National Institute of Allergy and Infectious Diseases Niaid
Fink, Doran L.
United States, Bethesda
National Institute of Allergy and Infectious Diseases Niaid
United States, Silver Spring
Food and Drug Administration
Kamgno, Joseph
Cameroon, Yaounde
Centre for Research on Filariasis and Other Tropical Diseases
Cameroon, Yaounde
Université de Yaoundé I
Herrick, Jesica A.
United States, Bethesda
National Institute of Allergy and Infectious Diseases Niaid
United States, Chicago
University of Illinois at Chicago
Nutman, B. Thomas
United States, Bethesda
National Institute of Allergy and Infectious Diseases Niaid
Statistics
Citations: 43
Authors: 5
Affiliations: 5
Identifiers
Doi:
10.1128/JCM.00525-14
ISSN:
00951137
e-ISSN:
1098660X
Research Areas
Genetics And Genomics
Infectious Diseases
Study Approach
Quantitative