A two-colour multiplexed lateral flow immunoassay system to differentially detect human malaria species on a single test line
Abstract Background Malaria continues to impose a tremendous burden in terms of global morbidity and mortality, yet even today, a large number of diagnoses are presumptive resulting in lack of or inappropriate treatment. Methods In this work, a two-colour lateral flow immunoassay (LFA) system was de...
Published in: | Malaria Journal |
---|---|
Main Authors: | , , , , , |
Format: | Article in Journal/Newspaper |
Language: | English |
Published: |
BMC
2019
|
Subjects: | |
Online Access: | https://doi.org/10.1186/s12936-019-2957-x https://doaj.org/article/5c6e0da4b5364b44a4fc3669abe87a42 |
id |
ftdoajarticles:oai:doaj.org/article:5c6e0da4b5364b44a4fc3669abe87a42 |
---|---|
record_format |
openpolar |
spelling |
ftdoajarticles:oai:doaj.org/article:5c6e0da4b5364b44a4fc3669abe87a42 2023-05-15T15:17:51+02:00 A two-colour multiplexed lateral flow immunoassay system to differentially detect human malaria species on a single test line Jinsu Kim Xiangkun Elvis Cao Julia L. Finkelstein Washington B. Cárdenas David Erickson Saurabh Mehta 2019-09-01T00:00:00Z https://doi.org/10.1186/s12936-019-2957-x https://doaj.org/article/5c6e0da4b5364b44a4fc3669abe87a42 EN eng BMC http://link.springer.com/article/10.1186/s12936-019-2957-x https://doaj.org/toc/1475-2875 doi:10.1186/s12936-019-2957-x 1475-2875 https://doaj.org/article/5c6e0da4b5364b44a4fc3669abe87a42 Malaria Journal, Vol 18, Iss 1, Pp 1-10 (2019) Malaria Diagnostics Screening Point of care Multiplex Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 article 2019 ftdoajarticles https://doi.org/10.1186/s12936-019-2957-x 2022-12-31T16:26:48Z Abstract Background Malaria continues to impose a tremendous burden in terms of global morbidity and mortality, yet even today, a large number of diagnoses are presumptive resulting in lack of or inappropriate treatment. Methods In this work, a two-colour lateral flow immunoassay (LFA) system was developed to identify infections by Plasmodium spp. and differentiate Plasmodium falciparum infection from the other three human malaria species (Plasmodium vivax, Plasmodium ovale, Plasmodium malariae). To achieve this goal, red and blue colours were encoded to two markers on a single test line of strips, for simultaneous detection of PfHRP2 (red), a marker specific for P. falciparum infection, and pLDH (blue), a pan-specific marker for infections by all species of Plasmodium. The assay performance was first optimized and evaluated with recombinant malarial proteins spiked in washing buffer at various concentrations from 0 to 1000 ng mL−1. The colour profiles developed on the single test line were discriminated and quantified: colour types corresponded to malaria protein species; colour intensities represented protein concentration levels. Results The limit of detection (the lowest concentrations of malaria antigens that can be distinguished from blank samples) and the limit of colour discrimination (the limit to differentiate pLDH from PfHRP2) were defined for the two-colour assay from the spiked buffer test, and the two limits were 31.2 ng mL−1 and 7.8 ng mL−1, respectively. To further validate the efficacy of the assay, 25 human whole blood frozen samples were tested and successfully validated against ELISA and microscopy results: 15 samples showed malaria negative; 5 samples showed P. falciparum positive; 5 samples showed P. falciparum negative, but contained other malaria species. Conclusions The assay provides a simple method to quickly identify and differentiate infection by different malarial parasites at the point-of-need and overcome the physical limitations of traditional LFAs, improving the multiplexing ... Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic Malaria Journal 18 1 |
institution |
Open Polar |
collection |
Directory of Open Access Journals: DOAJ Articles |
op_collection_id |
ftdoajarticles |
language |
English |
topic |
Malaria Diagnostics Screening Point of care Multiplex Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 |
spellingShingle |
Malaria Diagnostics Screening Point of care Multiplex Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 Jinsu Kim Xiangkun Elvis Cao Julia L. Finkelstein Washington B. Cárdenas David Erickson Saurabh Mehta A two-colour multiplexed lateral flow immunoassay system to differentially detect human malaria species on a single test line |
topic_facet |
Malaria Diagnostics Screening Point of care Multiplex Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 |
description |
Abstract Background Malaria continues to impose a tremendous burden in terms of global morbidity and mortality, yet even today, a large number of diagnoses are presumptive resulting in lack of or inappropriate treatment. Methods In this work, a two-colour lateral flow immunoassay (LFA) system was developed to identify infections by Plasmodium spp. and differentiate Plasmodium falciparum infection from the other three human malaria species (Plasmodium vivax, Plasmodium ovale, Plasmodium malariae). To achieve this goal, red and blue colours were encoded to two markers on a single test line of strips, for simultaneous detection of PfHRP2 (red), a marker specific for P. falciparum infection, and pLDH (blue), a pan-specific marker for infections by all species of Plasmodium. The assay performance was first optimized and evaluated with recombinant malarial proteins spiked in washing buffer at various concentrations from 0 to 1000 ng mL−1. The colour profiles developed on the single test line were discriminated and quantified: colour types corresponded to malaria protein species; colour intensities represented protein concentration levels. Results The limit of detection (the lowest concentrations of malaria antigens that can be distinguished from blank samples) and the limit of colour discrimination (the limit to differentiate pLDH from PfHRP2) were defined for the two-colour assay from the spiked buffer test, and the two limits were 31.2 ng mL−1 and 7.8 ng mL−1, respectively. To further validate the efficacy of the assay, 25 human whole blood frozen samples were tested and successfully validated against ELISA and microscopy results: 15 samples showed malaria negative; 5 samples showed P. falciparum positive; 5 samples showed P. falciparum negative, but contained other malaria species. Conclusions The assay provides a simple method to quickly identify and differentiate infection by different malarial parasites at the point-of-need and overcome the physical limitations of traditional LFAs, improving the multiplexing ... |
format |
Article in Journal/Newspaper |
author |
Jinsu Kim Xiangkun Elvis Cao Julia L. Finkelstein Washington B. Cárdenas David Erickson Saurabh Mehta |
author_facet |
Jinsu Kim Xiangkun Elvis Cao Julia L. Finkelstein Washington B. Cárdenas David Erickson Saurabh Mehta |
author_sort |
Jinsu Kim |
title |
A two-colour multiplexed lateral flow immunoassay system to differentially detect human malaria species on a single test line |
title_short |
A two-colour multiplexed lateral flow immunoassay system to differentially detect human malaria species on a single test line |
title_full |
A two-colour multiplexed lateral flow immunoassay system to differentially detect human malaria species on a single test line |
title_fullStr |
A two-colour multiplexed lateral flow immunoassay system to differentially detect human malaria species on a single test line |
title_full_unstemmed |
A two-colour multiplexed lateral flow immunoassay system to differentially detect human malaria species on a single test line |
title_sort |
two-colour multiplexed lateral flow immunoassay system to differentially detect human malaria species on a single test line |
publisher |
BMC |
publishDate |
2019 |
url |
https://doi.org/10.1186/s12936-019-2957-x https://doaj.org/article/5c6e0da4b5364b44a4fc3669abe87a42 |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
Malaria Journal, Vol 18, Iss 1, Pp 1-10 (2019) |
op_relation |
http://link.springer.com/article/10.1186/s12936-019-2957-x https://doaj.org/toc/1475-2875 doi:10.1186/s12936-019-2957-x 1475-2875 https://doaj.org/article/5c6e0da4b5364b44a4fc3669abe87a42 |
op_doi |
https://doi.org/10.1186/s12936-019-2957-x |
container_title |
Malaria Journal |
container_volume |
18 |
container_issue |
1 |
_version_ |
1766348106154115072 |