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Value of Routine Dengue Diagnostic Tests in Urine and Saliva Specimens
Dengue laboratory diagnosis is essentially based on detection of the virus, its components or antibodies directed against the virus in blood samples. Blood, however, may be difficult to draw in some patients, especially in children, and sampling during outbreak investigations or epidemiological stud...
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Published in: | PLoS neglected tropical diseases 2015-09, Vol.9 (9), p.e0004100-e0004100 |
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creator | Andries, Anne-Claire Duong, Veasna Ly, Sowath Cappelle, Julien Kim, Kim Srorn Lorn Try, Patrich Ros, Sopheaktra Ong, Sivuth Huy, Rekol Horwood, Paul Flamand, Marie Sakuntabhai, Anavaj Tarantola, Arnaud Buchy, Philippe |
description | Dengue laboratory diagnosis is essentially based on detection of the virus, its components or antibodies directed against the virus in blood samples. Blood, however, may be difficult to draw in some patients, especially in children, and sampling during outbreak investigations or epidemiological studies may face logistical challenges or limited compliance to invasive procedures from subjects. The aim of this study was to assess the possibility of using saliva and urine samples instead of blood for dengue diagnosis.
Serial plasma, urine and saliva samples were collected at several time-points between the day of admission to hospital until three months after the onset of fever in children with confirmed dengue disease. Quantitative RT-PCR, NS1 antigen capture and ELISA serology for anti-DENV antibody (IgG, IgM and IgA) detection were performed in parallel on the three body fluids. RT-PCR and NS1 tests demonstrated an overall sensitivity of 85.4%/63.4%, 41.6%/14.5% and 39%/28.3%, in plasma, urine and saliva specimens, respectively. When urine and saliva samples were collected at the same time-points and tested concurrently, the diagnostic sensitivity of RNA and NS1 detection assays was 69.1% and 34.4%, respectively. IgG/IgA detection assays had an overall sensitivity of 54.4%/37.4%, 38.5%/26.8% and 52.9%/28.6% in plasma, urine and saliva specimens, respectively. IgM were detected in 38.1% and 36% of the plasma and saliva samples but never in urine.
Although the performances of the different diagnostic methods were not as good in saliva and urine as in plasma specimens, the results obtained by qRT-PCR and by anti-DENV antibody ELISA could well justify the use of these two body fluids to detect dengue infection in situations when the collection of blood specimens is not possible. |
doi_str_mv | 10.1371/journal.pntd.0004100 |
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Serial plasma, urine and saliva samples were collected at several time-points between the day of admission to hospital until three months after the onset of fever in children with confirmed dengue disease. Quantitative RT-PCR, NS1 antigen capture and ELISA serology for anti-DENV antibody (IgG, IgM and IgA) detection were performed in parallel on the three body fluids. RT-PCR and NS1 tests demonstrated an overall sensitivity of 85.4%/63.4%, 41.6%/14.5% and 39%/28.3%, in plasma, urine and saliva specimens, respectively. When urine and saliva samples were collected at the same time-points and tested concurrently, the diagnostic sensitivity of RNA and NS1 detection assays was 69.1% and 34.4%, respectively. IgG/IgA detection assays had an overall sensitivity of 54.4%/37.4%, 38.5%/26.8% and 52.9%/28.6% in plasma, urine and saliva specimens, respectively. IgM were detected in 38.1% and 36% of the plasma and saliva samples but never in urine.
Although the performances of the different diagnostic methods were not as good in saliva and urine as in plasma specimens, the results obtained by qRT-PCR and by anti-DENV antibody ELISA could well justify the use of these two body fluids to detect dengue infection in situations when the collection of blood specimens is not possible.</description><identifier>ISSN: 1935-2735</identifier><identifier>ISSN: 1935-2727</identifier><identifier>EISSN: 1935-2735</identifier><identifier>DOI: 10.1371/journal.pntd.0004100</identifier><identifier>PMID: 26406240</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Adolescent ; Antibodies, Viral - analysis ; Antibodies, Viral - blood ; Antibodies, Viral - urine ; Antigens ; Body fluids ; Cambodia ; Child ; Child, Preschool ; Dengue ; Dengue - blood ; Dengue - diagnosis ; Dengue - epidemiology ; Dengue - urine ; Dengue fever ; Dengue Virus - genetics ; Dengue Virus - immunology ; Dengue Virus - isolation & purification ; Diagnosis ; Diagnostic Tests, Routine ; Disease ; Disease Progression ; Distribution ; Enzyme-Linked Immunosorbent Assay ; Epidemics ; Epidemiology ; Female ; Funding ; Genome, Viral ; Genomes ; Hospitals ; Humans ; Immunoglobulin Isotypes - analysis ; Immunoglobulin Isotypes - blood ; Immunoglobulin Isotypes - urine ; Infections ; Laboratories ; Life Sciences ; Male ; Microbiology and Parasitology ; Molecular diagnostic techniques ; Patients ; Physiological aspects ; Plasma ; Plasma - chemistry ; Plasma - immunology ; Plasma - virology ; Polymerase Chain Reaction ; RNA, Viral - isolation & purification ; Saliva ; Saliva - chemistry ; Saliva - immunology ; Saliva - virology ; Salivary glands ; secretions ; Studies ; Urine ; Urine - chemistry ; Urine - physiology ; Urine - virology ; Viral Nonstructural Proteins - analysis ; Viral Nonstructural Proteins - blood ; Viral Nonstructural Proteins - urine</subject><ispartof>PLoS neglected tropical diseases, 2015-09, Vol.9 (9), p.e0004100-e0004100</ispartof><rights>COPYRIGHT 2015 Public Library of Science</rights><rights>Attribution</rights><rights>2015 Andries et al 2015 Andries et al</rights><rights>2015 Public Library of Science. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited: Andries A-C, Duong V, Ly S, Cappelle J, Kim KS, Lorn Try P, et al. (2015) Value of Routine Dengue Diagnostic Tests in Urine and Saliva Specimens. 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A.</contributor><creatorcontrib>Andries, Anne-Claire</creatorcontrib><creatorcontrib>Duong, Veasna</creatorcontrib><creatorcontrib>Ly, Sowath</creatorcontrib><creatorcontrib>Cappelle, Julien</creatorcontrib><creatorcontrib>Kim, Kim Srorn</creatorcontrib><creatorcontrib>Lorn Try, Patrich</creatorcontrib><creatorcontrib>Ros, Sopheaktra</creatorcontrib><creatorcontrib>Ong, Sivuth</creatorcontrib><creatorcontrib>Huy, Rekol</creatorcontrib><creatorcontrib>Horwood, Paul</creatorcontrib><creatorcontrib>Flamand, Marie</creatorcontrib><creatorcontrib>Sakuntabhai, Anavaj</creatorcontrib><creatorcontrib>Tarantola, Arnaud</creatorcontrib><creatorcontrib>Buchy, Philippe</creatorcontrib><title>Value of Routine Dengue Diagnostic Tests in Urine and Saliva Specimens</title><title>PLoS neglected tropical diseases</title><addtitle>PLoS Negl Trop Dis</addtitle><description>Dengue laboratory diagnosis is essentially based on detection of the virus, its components or antibodies directed against the virus in blood samples. Blood, however, may be difficult to draw in some patients, especially in children, and sampling during outbreak investigations or epidemiological studies may face logistical challenges or limited compliance to invasive procedures from subjects. The aim of this study was to assess the possibility of using saliva and urine samples instead of blood for dengue diagnosis.
Serial plasma, urine and saliva samples were collected at several time-points between the day of admission to hospital until three months after the onset of fever in children with confirmed dengue disease. Quantitative RT-PCR, NS1 antigen capture and ELISA serology for anti-DENV antibody (IgG, IgM and IgA) detection were performed in parallel on the three body fluids. RT-PCR and NS1 tests demonstrated an overall sensitivity of 85.4%/63.4%, 41.6%/14.5% and 39%/28.3%, in plasma, urine and saliva specimens, respectively. When urine and saliva samples were collected at the same time-points and tested concurrently, the diagnostic sensitivity of RNA and NS1 detection assays was 69.1% and 34.4%, respectively. IgG/IgA detection assays had an overall sensitivity of 54.4%/37.4%, 38.5%/26.8% and 52.9%/28.6% in plasma, urine and saliva specimens, respectively. IgM were detected in 38.1% and 36% of the plasma and saliva samples but never in urine.
Although the performances of the different diagnostic methods were not as good in saliva and urine as in plasma specimens, the results obtained by qRT-PCR and by anti-DENV antibody ELISA could well justify the use of these two body fluids to detect dengue infection in situations when the collection of blood specimens is not possible.</description><subject>Adolescent</subject><subject>Antibodies, Viral - analysis</subject><subject>Antibodies, Viral - blood</subject><subject>Antibodies, Viral - urine</subject><subject>Antigens</subject><subject>Body fluids</subject><subject>Cambodia</subject><subject>Child</subject><subject>Child, Preschool</subject><subject>Dengue</subject><subject>Dengue - blood</subject><subject>Dengue - diagnosis</subject><subject>Dengue - epidemiology</subject><subject>Dengue - urine</subject><subject>Dengue fever</subject><subject>Dengue Virus - genetics</subject><subject>Dengue Virus - immunology</subject><subject>Dengue Virus - isolation & purification</subject><subject>Diagnosis</subject><subject>Diagnostic Tests, Routine</subject><subject>Disease</subject><subject>Disease Progression</subject><subject>Distribution</subject><subject>Enzyme-Linked Immunosorbent Assay</subject><subject>Epidemics</subject><subject>Epidemiology</subject><subject>Female</subject><subject>Funding</subject><subject>Genome, Viral</subject><subject>Genomes</subject><subject>Hospitals</subject><subject>Humans</subject><subject>Immunoglobulin Isotypes - analysis</subject><subject>Immunoglobulin Isotypes - blood</subject><subject>Immunoglobulin Isotypes - urine</subject><subject>Infections</subject><subject>Laboratories</subject><subject>Life Sciences</subject><subject>Male</subject><subject>Microbiology and Parasitology</subject><subject>Molecular diagnostic techniques</subject><subject>Patients</subject><subject>Physiological aspects</subject><subject>Plasma</subject><subject>Plasma - chemistry</subject><subject>Plasma - immunology</subject><subject>Plasma - virology</subject><subject>Polymerase Chain Reaction</subject><subject>RNA, Viral - isolation & purification</subject><subject>Saliva</subject><subject>Saliva - chemistry</subject><subject>Saliva - immunology</subject><subject>Saliva - virology</subject><subject>Salivary glands</subject><subject>secretions</subject><subject>Studies</subject><subject>Urine</subject><subject>Urine - chemistry</subject><subject>Urine - physiology</subject><subject>Urine - virology</subject><subject>Viral Nonstructural Proteins - analysis</subject><subject>Viral Nonstructural Proteins - blood</subject><subject>Viral Nonstructural Proteins - urine</subject><issn>1935-2735</issn><issn>1935-2727</issn><issn>1935-2735</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2015</creationdate><recordtype>article</recordtype><sourceid>DOA</sourceid><recordid>eNqNkluL2zAQhU1p6V7af1BaQ6H0JalutuSXQthLdyFQ6O72VSjyKFFQpFSyA_33lTfeJSl9KH6wGX_njGZ0iuIdRlNMOf6yDn30yk23vmunCCGGEXpRnOKGVhPCafXy4PukOEtpjVDVVAK_Lk5IzVBNGDotrn8q10MZTPkj9J31UF6CX-bKpVVLH1JndXkPqUul9eVDHADl2_JOObtT5d0WtN2AT2-KV0a5BG_H93nxcH11f3EzmX__dnsxm090XfNuYgSnDZBKCETwwhioGNRCI4MWWFGGCGlJZqDliukWgJqmJUQQpVmzUDWn58WHve_WhSTHFSSJOUEVxYSJTNzuiTaotdxGu1HxtwzKysdCiEupYp7KgWSaIaOpqBnGbEGMYpBbtkawilPcDF5fx279YgOtBt9F5Y5Mj_94u5LLsJOsEjTfUTaY7A1Wf8luZnO5VamDPkqEOc17EbuB_zw2jOFXn9cuNzZpcE55CP3jnLzmhBHyHygWiHPBWUY_7tGlylNbb0I-rB5wOWMUV5TWAmVq-g8qPy1srA4ejM31I8GnA8EKlOtWKbicouDTMcj2oI4hpQjmeRUYySHLTxcphyzLMctZ9v5w-8-ip_DSP-eO7ng</recordid><startdate>20150901</startdate><enddate>20150901</enddate><creator>Andries, Anne-Claire</creator><creator>Duong, Veasna</creator><creator>Ly, Sowath</creator><creator>Cappelle, Julien</creator><creator>Kim, Kim Srorn</creator><creator>Lorn Try, Patrich</creator><creator>Ros, Sopheaktra</creator><creator>Ong, Sivuth</creator><creator>Huy, Rekol</creator><creator>Horwood, Paul</creator><creator>Flamand, Marie</creator><creator>Sakuntabhai, Anavaj</creator><creator>Tarantola, Arnaud</creator><creator>Buchy, Philippe</creator><general>Public Library of Science</general><general>Public Library of Science (PLoS)</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope><scope>7T2</scope><scope>7U2</scope><scope>7U9</scope><scope>C1K</scope><scope>F1W</scope><scope>H94</scope><scope>H95</scope><scope>H97</scope><scope>L.G</scope><scope>1XC</scope><scope>VOOES</scope><scope>5PM</scope><scope>DOA</scope><orcidid>https://orcid.org/0000-0001-9099-6937</orcidid><orcidid>https://orcid.org/0000-0002-6946-7958</orcidid><orcidid>https://orcid.org/0000-0003-0353-1678</orcidid><orcidid>https://orcid.org/0000-0002-7104-7418</orcidid></search><sort><creationdate>20150901</creationdate><title>Value of Routine Dengue Diagnostic Tests in Urine and Saliva Specimens</title><author>Andries, Anne-Claire ; Duong, Veasna ; Ly, Sowath ; Cappelle, Julien ; Kim, Kim Srorn ; Lorn Try, Patrich ; Ros, Sopheaktra ; Ong, Sivuth ; Huy, Rekol ; Horwood, Paul ; Flamand, Marie ; Sakuntabhai, Anavaj ; Tarantola, Arnaud ; Buchy, Philippe</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c667t-f8739e2588021bffe54e68c0f0b1a34022d2f87ed7a4cdee3f9d2282ac49ba673</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2015</creationdate><topic>Adolescent</topic><topic>Antibodies, Viral - analysis</topic><topic>Antibodies, Viral - blood</topic><topic>Antibodies, Viral - urine</topic><topic>Antigens</topic><topic>Body fluids</topic><topic>Cambodia</topic><topic>Child</topic><topic>Child, Preschool</topic><topic>Dengue</topic><topic>Dengue - blood</topic><topic>Dengue - diagnosis</topic><topic>Dengue - epidemiology</topic><topic>Dengue - urine</topic><topic>Dengue fever</topic><topic>Dengue Virus - genetics</topic><topic>Dengue Virus - immunology</topic><topic>Dengue Virus - isolation & purification</topic><topic>Diagnosis</topic><topic>Diagnostic Tests, Routine</topic><topic>Disease</topic><topic>Disease Progression</topic><topic>Distribution</topic><topic>Enzyme-Linked Immunosorbent Assay</topic><topic>Epidemics</topic><topic>Epidemiology</topic><topic>Female</topic><topic>Funding</topic><topic>Genome, Viral</topic><topic>Genomes</topic><topic>Hospitals</topic><topic>Humans</topic><topic>Immunoglobulin Isotypes - analysis</topic><topic>Immunoglobulin Isotypes - blood</topic><topic>Immunoglobulin Isotypes - urine</topic><topic>Infections</topic><topic>Laboratories</topic><topic>Life Sciences</topic><topic>Male</topic><topic>Microbiology and Parasitology</topic><topic>Molecular diagnostic techniques</topic><topic>Patients</topic><topic>Physiological aspects</topic><topic>Plasma</topic><topic>Plasma - chemistry</topic><topic>Plasma - immunology</topic><topic>Plasma - virology</topic><topic>Polymerase Chain Reaction</topic><topic>RNA, Viral - isolation & purification</topic><topic>Saliva</topic><topic>Saliva - chemistry</topic><topic>Saliva - immunology</topic><topic>Saliva - virology</topic><topic>Salivary glands</topic><topic>secretions</topic><topic>Studies</topic><topic>Urine</topic><topic>Urine - chemistry</topic><topic>Urine - physiology</topic><topic>Urine - virology</topic><topic>Viral Nonstructural Proteins - analysis</topic><topic>Viral Nonstructural Proteins - blood</topic><topic>Viral Nonstructural Proteins - urine</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Andries, Anne-Claire</creatorcontrib><creatorcontrib>Duong, Veasna</creatorcontrib><creatorcontrib>Ly, Sowath</creatorcontrib><creatorcontrib>Cappelle, Julien</creatorcontrib><creatorcontrib>Kim, Kim Srorn</creatorcontrib><creatorcontrib>Lorn Try, Patrich</creatorcontrib><creatorcontrib>Ros, Sopheaktra</creatorcontrib><creatorcontrib>Ong, Sivuth</creatorcontrib><creatorcontrib>Huy, Rekol</creatorcontrib><creatorcontrib>Horwood, Paul</creatorcontrib><creatorcontrib>Flamand, Marie</creatorcontrib><creatorcontrib>Sakuntabhai, Anavaj</creatorcontrib><creatorcontrib>Tarantola, Arnaud</creatorcontrib><creatorcontrib>Buchy, Philippe</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>Health and Safety Science Abstracts (Full archive)</collection><collection>Safety Science and Risk</collection><collection>Virology and AIDS Abstracts</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ASFA: Aquatic Sciences and Fisheries Abstracts</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Aquatic Science & Fisheries Abstracts (ASFA) 1: Biological Sciences & Living Resources</collection><collection>Aquatic Science & Fisheries Abstracts (ASFA) 3: Aquatic Pollution & Environmental Quality</collection><collection>Aquatic Science & Fisheries Abstracts (ASFA) Professional</collection><collection>Hyper Article en Ligne (HAL)</collection><collection>Hyper Article en Ligne (HAL) (Open Access)</collection><collection>PubMed Central (Full Participant titles)</collection><collection>Directory of Open Access Journals</collection><jtitle>PLoS neglected tropical diseases</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Andries, Anne-Claire</au><au>Duong, Veasna</au><au>Ly, Sowath</au><au>Cappelle, Julien</au><au>Kim, Kim Srorn</au><au>Lorn Try, Patrich</au><au>Ros, Sopheaktra</au><au>Ong, Sivuth</au><au>Huy, Rekol</au><au>Horwood, Paul</au><au>Flamand, Marie</au><au>Sakuntabhai, Anavaj</au><au>Tarantola, Arnaud</au><au>Buchy, Philippe</au><au>Marques, Ernesto T. 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The aim of this study was to assess the possibility of using saliva and urine samples instead of blood for dengue diagnosis.
Serial plasma, urine and saliva samples were collected at several time-points between the day of admission to hospital until three months after the onset of fever in children with confirmed dengue disease. Quantitative RT-PCR, NS1 antigen capture and ELISA serology for anti-DENV antibody (IgG, IgM and IgA) detection were performed in parallel on the three body fluids. RT-PCR and NS1 tests demonstrated an overall sensitivity of 85.4%/63.4%, 41.6%/14.5% and 39%/28.3%, in plasma, urine and saliva specimens, respectively. When urine and saliva samples were collected at the same time-points and tested concurrently, the diagnostic sensitivity of RNA and NS1 detection assays was 69.1% and 34.4%, respectively. IgG/IgA detection assays had an overall sensitivity of 54.4%/37.4%, 38.5%/26.8% and 52.9%/28.6% in plasma, urine and saliva specimens, respectively. IgM were detected in 38.1% and 36% of the plasma and saliva samples but never in urine.
Although the performances of the different diagnostic methods were not as good in saliva and urine as in plasma specimens, the results obtained by qRT-PCR and by anti-DENV antibody ELISA could well justify the use of these two body fluids to detect dengue infection in situations when the collection of blood specimens is not possible.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>26406240</pmid><doi>10.1371/journal.pntd.0004100</doi><orcidid>https://orcid.org/0000-0001-9099-6937</orcidid><orcidid>https://orcid.org/0000-0002-6946-7958</orcidid><orcidid>https://orcid.org/0000-0003-0353-1678</orcidid><orcidid>https://orcid.org/0000-0002-7104-7418</orcidid><oa>free_for_read</oa></addata></record> |
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recordid | cdi_plos_journals_1720531248 |
source | Access via ProQuest (Open Access); PubMed Central |
subjects | Adolescent Antibodies, Viral - analysis Antibodies, Viral - blood Antibodies, Viral - urine Antigens Body fluids Cambodia Child Child, Preschool Dengue Dengue - blood Dengue - diagnosis Dengue - epidemiology Dengue - urine Dengue fever Dengue Virus - genetics Dengue Virus - immunology Dengue Virus - isolation & purification Diagnosis Diagnostic Tests, Routine Disease Disease Progression Distribution Enzyme-Linked Immunosorbent Assay Epidemics Epidemiology Female Funding Genome, Viral Genomes Hospitals Humans Immunoglobulin Isotypes - analysis Immunoglobulin Isotypes - blood Immunoglobulin Isotypes - urine Infections Laboratories Life Sciences Male Microbiology and Parasitology Molecular diagnostic techniques Patients Physiological aspects Plasma Plasma - chemistry Plasma - immunology Plasma - virology Polymerase Chain Reaction RNA, Viral - isolation & purification Saliva Saliva - chemistry Saliva - immunology Saliva - virology Salivary glands secretions Studies Urine Urine - chemistry Urine - physiology Urine - virology Viral Nonstructural Proteins - analysis Viral Nonstructural Proteins - blood Viral Nonstructural Proteins - urine |
title | Value of Routine Dengue Diagnostic Tests in Urine and Saliva Specimens |
url | http://sfxeu10.hosted.exlibrisgroup.com/loughborough?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-25T08%3A15%3A36IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-gale_plos_&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Value%20of%20Routine%20Dengue%20Diagnostic%20Tests%20in%20Urine%20and%20Saliva%20Specimens&rft.jtitle=PLoS%20neglected%20tropical%20diseases&rft.au=Andries,%20Anne-Claire&rft.date=2015-09-01&rft.volume=9&rft.issue=9&rft.spage=e0004100&rft.epage=e0004100&rft.pages=e0004100-e0004100&rft.issn=1935-2735&rft.eissn=1935-2735&rft_id=info:doi/10.1371/journal.pntd.0004100&rft_dat=%3Cgale_plos_%3EA431533680%3C/gale_plos_%3E%3Cgrp_id%3Ecdi_FETCH-LOGICAL-c667t-f8739e2588021bffe54e68c0f0b1a34022d2f87ed7a4cdee3f9d2282ac49ba673%3C/grp_id%3E%3Coa%3E%3C/oa%3E%3Curl%3E%3C/url%3E&rft_id=info:oai/&rft_pqid=1718077874&rft_id=info:pmid/26406240&rft_galeid=A431533680&rfr_iscdi=true |