Quantitative detection of <it>Pf</it>HRP2 in saliva of malaria patients in the Philippines

<p>Abstract</p> <p>Background</p> <p>Malaria is a global health priority with a heavy burden of fatality and morbidity. Improvements in field diagnostics are needed to support the agenda for malaria elimination. Saliva has shown significant potential for use in non-inva...

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Main Authors: Fung Andrew O, Damoiseaux Robert, Grundeen Sarah, Panes Jonnas L, Horton Daniel H, Judy Jack W, Moore Theodore B
Format: Article
Language:English
Published: BMC 2012-05-01
Series:Malaria Journal
Online Access:http://www.malariajournal.com/content/11/1/175
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author Fung Andrew O
Damoiseaux Robert
Grundeen Sarah
Panes Jonnas L
Horton Daniel H
Judy Jack W
Moore Theodore B
author_facet Fung Andrew O
Damoiseaux Robert
Grundeen Sarah
Panes Jonnas L
Horton Daniel H
Judy Jack W
Moore Theodore B
author_sort Fung Andrew O
collection DOAJ
description <p>Abstract</p> <p>Background</p> <p>Malaria is a global health priority with a heavy burden of fatality and morbidity. Improvements in field diagnostics are needed to support the agenda for malaria elimination. Saliva has shown significant potential for use in non-invasive diagnostics, but the development of off-the-shelf saliva diagnostic kits requires best practices for sample preparation and quantitative insight on the availability of biomarkers and the dynamics of immunoassay in saliva. This pilot study measured the levels of the <it>Pf</it>HRP2 in patient saliva to inform the development of salivary diagnostic tests for malaria.</p> <p>Methods</p> <p>Matched samples of blood and saliva were collected between January and May, 2011 from eight patients at Palawan Baptist Hospital in Roxas, Palawan, Philippines. Parasite density was determined from thick-film blood smears. Concentrations of <it>Pf</it>HRP2 in saliva of malaria-positive patients were measured using a custom chemiluminescent ELISA in microtitre plates. Sixteen negative-control patients were enrolled at UCLA. A substantive difference between this protocol and previous related studies was that saliva samples were stabilized with protease inhibitors.</p> <p>Results</p> <p>Of the eight patients with microscopically confirmed <it>P. falciparum</it> malaria, seven tested positive for <it>Pf</it>HRP2 in the blood using rapid diagnostic test kits, and all tested positive for <it>Pf</it>HRP2 in saliva. All negative-control samples tested negative for salivary <it>Pf</it>HRP2. On a binary-decision basis, the ELISA agreed with microscopy with 100 % sensitivity and 100 % specificity. Salivary levels of <it>Pf</it>HRP2 ranged from 17 to 1,167 pg/mL in the malaria-positive group.</p> <p>Conclusion</p> <p>Saliva is a promising diagnostic fluid for malaria when protein degradation and matrix effects are mitigated. Systematic quantitation of other malaria biomarkers in saliva would identify those with the best clinical relevance and suitability for off-the-shelf diagnostic kits.</p>
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spelling doaj.art-1ddf840778da4496b67a3caad5e910f42022-12-21T23:22:07ZengBMCMalaria Journal1475-28752012-05-0111117510.1186/1475-2875-11-175Quantitative detection of <it>Pf</it>HRP2 in saliva of malaria patients in the PhilippinesFung Andrew ODamoiseaux RobertGrundeen SarahPanes Jonnas LHorton Daniel HJudy Jack WMoore Theodore B<p>Abstract</p> <p>Background</p> <p>Malaria is a global health priority with a heavy burden of fatality and morbidity. Improvements in field diagnostics are needed to support the agenda for malaria elimination. Saliva has shown significant potential for use in non-invasive diagnostics, but the development of off-the-shelf saliva diagnostic kits requires best practices for sample preparation and quantitative insight on the availability of biomarkers and the dynamics of immunoassay in saliva. This pilot study measured the levels of the <it>Pf</it>HRP2 in patient saliva to inform the development of salivary diagnostic tests for malaria.</p> <p>Methods</p> <p>Matched samples of blood and saliva were collected between January and May, 2011 from eight patients at Palawan Baptist Hospital in Roxas, Palawan, Philippines. Parasite density was determined from thick-film blood smears. Concentrations of <it>Pf</it>HRP2 in saliva of malaria-positive patients were measured using a custom chemiluminescent ELISA in microtitre plates. Sixteen negative-control patients were enrolled at UCLA. A substantive difference between this protocol and previous related studies was that saliva samples were stabilized with protease inhibitors.</p> <p>Results</p> <p>Of the eight patients with microscopically confirmed <it>P. falciparum</it> malaria, seven tested positive for <it>Pf</it>HRP2 in the blood using rapid diagnostic test kits, and all tested positive for <it>Pf</it>HRP2 in saliva. All negative-control samples tested negative for salivary <it>Pf</it>HRP2. On a binary-decision basis, the ELISA agreed with microscopy with 100 % sensitivity and 100 % specificity. Salivary levels of <it>Pf</it>HRP2 ranged from 17 to 1,167 pg/mL in the malaria-positive group.</p> <p>Conclusion</p> <p>Saliva is a promising diagnostic fluid for malaria when protein degradation and matrix effects are mitigated. Systematic quantitation of other malaria biomarkers in saliva would identify those with the best clinical relevance and suitability for off-the-shelf diagnostic kits.</p>http://www.malariajournal.com/content/11/1/175
spellingShingle Fung Andrew O
Damoiseaux Robert
Grundeen Sarah
Panes Jonnas L
Horton Daniel H
Judy Jack W
Moore Theodore B
Quantitative detection of <it>Pf</it>HRP2 in saliva of malaria patients in the Philippines
Malaria Journal
title Quantitative detection of <it>Pf</it>HRP2 in saliva of malaria patients in the Philippines
title_full Quantitative detection of <it>Pf</it>HRP2 in saliva of malaria patients in the Philippines
title_fullStr Quantitative detection of <it>Pf</it>HRP2 in saliva of malaria patients in the Philippines
title_full_unstemmed Quantitative detection of <it>Pf</it>HRP2 in saliva of malaria patients in the Philippines
title_short Quantitative detection of <it>Pf</it>HRP2 in saliva of malaria patients in the Philippines
title_sort quantitative detection of it pf it hrp2 in saliva of malaria patients in the philippines
url http://www.malariajournal.com/content/11/1/175
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