Parasitological Confirmation and Analysis of Leishmania Diversity in Asymptomatic and Subclinical Infection following Resolution of Cutaneous Leishmaniasis
Publication Date
December 11, 2015
Journal
PLOS Neglected Tropical Diseases
Authors
Mariana Rosales Chilama, Rafael E. Gongora, Liliana Valderrama, Jimena Jojoa, et al
Volume
9
Issue
12
Pages
e0004273
DOI
https://dx.plos.org/10.1371/journal.pntd.0004273
Publisher URL
http://journals.plos.org/plosntds/article?id=10.1371%2Fjournal.pntd.0004273
PubMed
http://www.ncbi.nlm.nih.gov/pubmed/26659114
PubMed Central
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4684356
Europe PMC
http://europepmc.org/abstract/MED/26659114
Web of Science
000368345100047
Scopus
84953230705
Mendeley
http://www.mendeley.com/research/parasitological-confirmation-analysis-leishmania-diversity-asymptomatic-subclinical-infection-follow
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Mendeley | Further Information

{"title"=>"Parasitological Confirmation and Analysis of Leishmania Diversity in Asymptomatic and Subclinical Infection following Resolution of Cutaneous Leishmaniasis", "type"=>"journal", "authors"=>[{"first_name"=>"Mariana", "last_name"=>"Rosales-Chilama", "scopus_author_id"=>"56650982400"}, {"first_name"=>"Rafael E.", "last_name"=>"Gongora", "scopus_author_id"=>"7003448085"}, {"first_name"=>"Liliana", "last_name"=>"Valderrama", "scopus_author_id"=>"6701830781"}, {"first_name"=>"Jimena", "last_name"=>"Jojoa", "scopus_author_id"=>"55981973900"}, {"first_name"=>"Neal", "last_name"=>"Alexander", "scopus_author_id"=>"7203015251"}, {"first_name"=>"Luisa C.", "last_name"=>"Rubiano", "scopus_author_id"=>"54928237600"}, {"first_name"=>"Alexandra", "last_name"=>"Cossio", "scopus_author_id"=>"55820774800"}, {"first_name"=>"Emily R.", "last_name"=>"Adams", "scopus_author_id"=>"15076919600"}, {"first_name"=>"Nancy G.", "last_name"=>"Saravia", "scopus_author_id"=>"7004212910"}, {"first_name"=>"María Adelaida", "last_name"=>"Gomez", "scopus_author_id"=>"10143098900"}], "year"=>2015, "source"=>"PLoS Neglected Tropical Diseases", "identifiers"=>{"issn"=>"19352735", "scopus"=>"2-s2.0-84953230705", "sgr"=>"84953230705", "pui"=>"607461917", "pmid"=>"26659114", "doi"=>"10.1371/journal.pntd.0004273"}, "id"=>"5e125593-dc78-38ae-b7a7-d06864653d24", "abstract"=>"BACKGROUND: The contribution of individuals with subclinical infection to the transmission and endemicity of cutaneous leishmaniasis (CL) is unknown. Immunological evidence of exposure to Leishmania in residents of endemic areas has been the basis for defining the human population with asymptomatic infection. However, parasitological confirmation of subclinical infection is lacking.\\n\\nMETHODS: We investigated the presence and viability of Leishmania in blood and non-invasive mucosal tissue samples from individuals with immunological evidence of subclinical infection in endemic areas for CL caused by Leishmania (Viannia) in Colombia. Detection of Leishmania kDNA was conducted by PCR-Southern Blot, and parasite viability was confirmed by amplification of parasite 7SLRNA gene transcripts. A molecular tool for genetic diversity analysis of parasite populations causing persistent subclinical infection based on PCR amplification and sequence analysis of an 82bp region between kDNA conserved blocks 1 and 2 was developed.\\n\\nPRINCIPAL FINDINGS: Persistent Leishmania infection was demonstrated in 40% (46 of 114) of leishmanin skin test (LST) positive individuals without active disease; parasite viability was established in 59% of these (27 of 46; 24% of total). Parasite burden quantified from circulating blood monocytes, nasal, conjunctival or tonsil mucosal swab samples was comparable, and ranged between 0.2 to 22 parasites per reaction. kDNA sequences were obtained from samples from 2 individuals with asymptomatic infection and from 26 with history of CL, allowing genetic distance analysis that revealed diversity among sequences and clustering within the L. (Viannia) subgenus.\\n\\nCONCLUSIONS: Our results provide parasitological confirmation of persistent infection among residents of endemic areas of L. (Viannia) transmission who have experienced asymptomatic infection or recovered from CL, revealing a reservoir of infection that potentially contributes to the endemicity and transmission of disease. kDNA genotyping establishes proof-of-principle of the feasibility of genetic diversity analysis in previously inaccessible and unexplored parasite populations in subclinically infected individuals.", "link"=>"http://www.mendeley.com/research/parasitological-confirmation-analysis-leishmania-diversity-asymptomatic-subclinical-infection-follow", "reader_count"=>45, "reader_count_by_academic_status"=>{"Unspecified"=>3, "Professor > Associate Professor"=>4, "Researcher"=>8, "Student > Doctoral Student"=>2, "Student > Ph. D. Student"=>6, "Student > Postgraduate"=>2, "Student > Master"=>11, "Other"=>3, "Student > Bachelor"=>4, "Professor"=>2}, "reader_count_by_user_role"=>{"Unspecified"=>3, "Professor > Associate Professor"=>4, "Researcher"=>8, "Student > Doctoral Student"=>2, "Student > Ph. D. Student"=>6, "Student > Postgraduate"=>2, "Student > Master"=>11, "Other"=>3, "Student > Bachelor"=>4, "Professor"=>2}, "reader_count_by_subject_area"=>{"Unspecified"=>3, "Biochemistry, Genetics and Molecular Biology"=>6, "Nursing and Health Professions"=>1, "Agricultural and Biological Sciences"=>19, "Medicine and Dentistry"=>9, "Veterinary Science and Veterinary Medicine"=>2, "Immunology and Microbiology"=>5}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>9}, "Immunology and Microbiology"=>{"Immunology and Microbiology"=>5}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>19}, "Nursing and Health Professions"=>{"Nursing and Health Professions"=>1}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>6}, "Unspecified"=>{"Unspecified"=>3}, "Veterinary Science and Veterinary Medicine"=>{"Veterinary Science and Veterinary Medicine"=>2}}, "reader_count_by_country"=>{"Sri Lanka"=>1, "Brazil"=>2, "Peru"=>1}, "group_count"=>5}

Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/2607380"], "description"=>"<p>Schematic representation of study phases and study population.</p>", "links"=>[], "tags"=>["Cutaneous Leishmaniasis BackgroundThe contribution", "leishmanin skin test", "parasitological confirmation", "parasite viability", "subclinical infection", "pcr", "individual", "Diversity analysis", "lst", "kdna", "mucosal tissue samples", "82 bp region", "developed.Principal FindingsPersistent Leishmania infection", "parasite populations", "cl", "parasite 7 SLRNA gene transcripts"], "article_id"=>1623845, "categories"=>["Biological Sciences", "Ecology"], "users"=>["Mariana Rosales-Chilama", "Rafael E. Gongora", "Liliana Valderrama", "Jimena Jojoa", "Neal Alexander", "Luisa C. Rubiano", "Alexandra Cossio", "Emily R. Adams", "Nancy G. Saravia", "María Adelaida Gómez"], "doi"=>"https://dx.doi.org/10.1371/journal.pntd.0004273.g001", "stats"=>{"downloads"=>0, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Schematic_representation_of_study_phases_and_study_population_/1623845", "title"=>"Schematic representation of study phases and study population.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-12-17 08:09:02"}
  • {"files"=>["https://ndownloader.figshare.com/files/2607381"], "description"=>"<p><i>Leishmania</i> minicircle kDNA was detected by PCR amplification and southern blot. Lanes 1–6: Tonsil swab samples; Lanes 7–12: conjunctival swab samples; Lanes 13–18: nasal mucosa swab samples; Lanes 19–24: blood monocytes. HI: History of CL, H: Healthy, A: Asymptomatic. The arrow denotes the 700bp band corresponding to the full length amplification product of <i>Leishmania</i> minicircle kDNA. The lower band represents an unspecific kDNA PCR product as confirmed by sequencing.</p>", "links"=>[], "tags"=>["Cutaneous Leishmaniasis BackgroundThe contribution", "leishmanin skin test", "parasitological confirmation", "parasite viability", "subclinical infection", "pcr", "individual", "Diversity analysis", "lst", "kdna", "mucosal tissue samples", "82 bp region", "developed.Principal FindingsPersistent Leishmania infection", "parasite populations", "cl", "parasite 7 SLRNA gene transcripts"], "article_id"=>1623846, "categories"=>["Biological Sciences", "Ecology"], "users"=>["Mariana Rosales-Chilama", "Rafael E. Gongora", "Liliana Valderrama", "Jimena Jojoa", "Neal Alexander", "Luisa C. Rubiano", "Alexandra Cossio", "Emily R. Adams", "Nancy G. Saravia", "María Adelaida Gómez"], "doi"=>"https://dx.doi.org/10.1371/journal.pntd.0004273.g002", "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Detection_of_Leishmania_kDNA_in_samples_from_individuals_with_asymptomatic_infection_who_share_a_household_with_a_historic_symptomatic_case_of_CL_/1623846", "title"=>"Detection of <i>Leishmania</i> kDNA in samples from individuals with asymptomatic infection who share a household with a historic symptomatic case of CL.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-12-17 08:09:09"}
  • {"files"=>["https://ndownloader.figshare.com/files/2607382"], "description"=>"<p>Absolute quantification of <i>Leishmania</i><b>(A)</b> and human nucleated cells <b>(B)</b> by qRT-PCR amplification of 7SLRNA and TBP transcripts, respectively, from blood monocytes (n = 33) and mucosal swab samples (n = 37) obtained from tonsil (triangles), conjunctiva (squares) and nasal mucosa (circles). Data are also shown as relative parasite burden normalized to the number of human cells per sample <b>(C).</b> Filled black figures represent LST+ individuals with clinical history of CL; open figures, LST+ individuals with asymptomatic infection; and grey figures, healthy LST- individuals from endemic area for CL. <i>p</i> values obtained from two tailed Mann-Whitney tests are shown.</p>", "links"=>[], "tags"=>["Cutaneous Leishmaniasis BackgroundThe contribution", "leishmanin skin test", "parasitological confirmation", "parasite viability", "subclinical infection", "pcr", "individual", "Diversity analysis", "lst", "kdna", "mucosal tissue samples", "82 bp region", "developed.Principal FindingsPersistent Leishmania infection", "parasite populations", "cl", "parasite 7 SLRNA gene transcripts"], "article_id"=>1623847, "categories"=>["Biological Sciences", "Ecology"], "users"=>["Mariana Rosales-Chilama", "Rafael E. Gongora", "Liliana Valderrama", "Jimena Jojoa", "Neal Alexander", "Luisa C. Rubiano", "Alexandra Cossio", "Emily R. Adams", "Nancy G. Saravia", "María Adelaida Gómez"], "doi"=>"https://dx.doi.org/10.1371/journal.pntd.0004273.g003", "stats"=>{"downloads"=>3, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Leishmania_viability_and_parasite_burden_in_kDNA_positive_samples_/1623847", "title"=>"<i>Leishmania</i> viability and parasite burden in kDNA positive samples.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-12-17 08:08:59"}
  • {"files"=>["https://ndownloader.figshare.com/files/2607383"], "description"=>"<p>Nested PCR amplification products from serial dilution curves of DNA from <i>L</i>. <i>V</i>. <i>panamensis</i> promastigotes <b>(A)</b> and human PBMCs <b>(B).</b> The upper panel shows the first amplification product of 700 bp obtained with LVp1 primer sets (external primers) and the bottom panel shows the 180 bp product of the nested PCR obtained with LVp1-LVp5 primer sets (internal primers). Nested PCR amplification products from reference strains of <i>L</i>.<i>V</i>. <i>panamensis</i> (L.p), <i>L</i>.<i>V</i>. <i>guyanensis</i> (L.g), <i>L</i>.<i>V</i>. <i>braziliensis</i> (L.b), <i>L</i>. <i>infantum</i> (L. inf), <i>L</i>. <i>amazonensis</i> (L.ama), <i>L</i>. <i>mexicana</i> (L. mex) and <i>T</i>. <i>cruzi</i><b>(C).</b> C-, negative control—water; PBMCs, DNA from uninfected human PBMCs.</p>", "links"=>[], "tags"=>["Cutaneous Leishmaniasis BackgroundThe contribution", "leishmanin skin test", "parasitological confirmation", "parasite viability", "subclinical infection", "pcr", "individual", "Diversity analysis", "lst", "kdna", "mucosal tissue samples", "82 bp region", "developed.Principal FindingsPersistent Leishmania infection", "parasite populations", "cl", "parasite 7 SLRNA gene transcripts"], "article_id"=>1623848, "categories"=>["Biological Sciences", "Ecology"], "users"=>["Mariana Rosales-Chilama", "Rafael E. Gongora", "Liliana Valderrama", "Jimena Jojoa", "Neal Alexander", "Luisa C. Rubiano", "Alexandra Cossio", "Emily R. Adams", "Nancy G. Saravia", "María Adelaida Gómez"], "doi"=>"https://dx.doi.org/10.1371/journal.pntd.0004273.g004", "stats"=>{"downloads"=>0, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Sensitivity_and_specificity_of_the_nested_PCR_for_Leishmania_minicircle_kDNA_/1623848", "title"=>"Sensitivity and specificity of the nested PCR for <i>Leishmania</i> minicircle kDNA.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-12-17 08:09:06"}
  • {"files"=>["https://ndownloader.figshare.com/files/2607384"], "description"=>"<p>An 82bp sequence spanning conserved blocks 1 and 2 from the <i>Leishmania</i> minicircle kDNA was used for multiple sequence alignment. Sequences from five different strains were analyzed for each <i>Leishmania</i> species, and from two <i>T</i>. <i>cruzi</i> strains. Strain details are summarized in <a href=\"http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0004273#pntd.0004273.s004\" target=\"_blank\">S1</a> and <a href=\"http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0004273#pntd.0004273.s005\" target=\"_blank\">S2</a> Tables. CB: Conserved Block.</p>", "links"=>[], "tags"=>["Cutaneous Leishmaniasis BackgroundThe contribution", "leishmanin skin test", "parasitological confirmation", "parasite viability", "subclinical infection", "pcr", "individual", "Diversity analysis", "lst", "kdna", "mucosal tissue samples", "82 bp region", "developed.Principal FindingsPersistent Leishmania infection", "parasite populations", "cl", "parasite 7 SLRNA gene transcripts"], "article_id"=>1623849, "categories"=>["Biological Sciences", "Ecology"], "users"=>["Mariana Rosales-Chilama", "Rafael E. Gongora", "Liliana Valderrama", "Jimena Jojoa", "Neal Alexander", "Luisa C. Rubiano", "Alexandra Cossio", "Emily R. Adams", "Nancy G. Saravia", "María Adelaida Gómez"], "doi"=>"https://dx.doi.org/10.1371/journal.pntd.0004273.g005", "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Analysis_of_Leishmania_genetic_diversity_/1623849", "title"=>"Analysis of <i>Leishmania</i> genetic diversity.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-12-17 08:09:02"}
  • {"files"=>["https://ndownloader.figshare.com/files/2607385"], "description"=>"<p>UPGMA trees of distances calculated from MLMT data <b>(A)</b> and conserved block minicircle kDNA genotyping <b>(B)</b> from a panel of 34 <i>L</i>. <i>V</i>. <i>panamensis</i> and 9 <i>L</i>. <i>V</i>. <i>guyanensis</i> clinical strains. Figure labels depict the geographical origin of strains within the Colombian territory. Shadowed codes denote groups of strains isolated from foci of transmission during outbreaks of infection.</p>", "links"=>[], "tags"=>["Cutaneous Leishmaniasis BackgroundThe contribution", "leishmanin skin test", "parasitological confirmation", "parasite viability", "subclinical infection", "pcr", "individual", "Diversity analysis", "lst", "kdna", "mucosal tissue samples", "82 bp region", "developed.Principal FindingsPersistent Leishmania infection", "parasite populations", "cl", "parasite 7 SLRNA gene transcripts"], "article_id"=>1623850, "categories"=>["Biological Sciences", "Ecology"], "users"=>["Mariana Rosales-Chilama", "Rafael E. Gongora", "Liliana Valderrama", "Jimena Jojoa", "Neal Alexander", "Luisa C. Rubiano", "Alexandra Cossio", "Emily R. Adams", "Nancy G. Saravia", "María Adelaida Gómez"], "doi"=>"https://dx.doi.org/10.1371/journal.pntd.0004273.g006", "stats"=>{"downloads"=>0, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Genetic_diversity_analysis_using_minicircle_kDNA_genotyping_allows_clustering_of_strains_isolated_within_foci_of_transmission_during_outbreaks_of_CL_/1623850", "title"=>"Genetic diversity analysis using minicircle kDNA genotyping allows clustering of strains isolated within foci of transmission during outbreaks of CL.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-12-17 08:08:59"}
  • {"files"=>["https://ndownloader.figshare.com/files/2607386"], "description"=>"<p>Sequences from the 82bp sequence spanning conserved blocks 1 and 2 from <i>Leishmania</i> minicircle kDNA were analyzed. Shown is a UPGMA tree of the genetic distances of sequences from clinical samples obtained from study participants with subclinical infection (blue and red triangles) and strains isolated from study participants with active disease (yellow circles), alongside a panel of 34 <i>L</i>. (<i>V</i>) <i>panamensis</i>, 10 <i>L</i>. (<i>V</i>.) <i>braziliensis</i>, and 9 <i>L</i>. (<i>V</i>) <i>guyanensis</i> clinical strains. Minicircle kDNA sequences retrieved from NCBI Genbank from <i>9 L</i>. <i>donovani</i> and 8 <i>L</i>. <i>infantum</i> strains were used as outgroups.</p>", "links"=>[], "tags"=>["Cutaneous Leishmaniasis BackgroundThe contribution", "leishmanin skin test", "parasitological confirmation", "parasite viability", "subclinical infection", "pcr", "individual", "Diversity analysis", "lst", "kdna", "mucosal tissue samples", "82 bp region", "developed.Principal FindingsPersistent Leishmania infection", "parasite populations", "cl", "parasite 7 SLRNA gene transcripts"], "article_id"=>1623851, "categories"=>["Biological Sciences", "Ecology"], "users"=>["Mariana Rosales-Chilama", "Rafael E. Gongora", "Liliana Valderrama", "Jimena Jojoa", "Neal Alexander", "Luisa C. Rubiano", "Alexandra Cossio", "Emily R. Adams", "Nancy G. Saravia", "María Adelaida Gómez"], "doi"=>"https://dx.doi.org/10.1371/journal.pntd.0004273.g007", "stats"=>{"downloads"=>0, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Analysis_of_Leishmania_genetic_diversity_in_individuals_with_infection_in_the_absence_of_disease_/1623851", "title"=>"Analysis of <i>Leishmania</i> genetic diversity in individuals with infection in the absence of disease.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-12-17 08:08:59"}
  • {"files"=>["https://ndownloader.figshare.com/files/2607387", "https://ndownloader.figshare.com/files/2607388", "https://ndownloader.figshare.com/files/2607389", "https://ndownloader.figshare.com/files/2607390", "https://ndownloader.figshare.com/files/2607391", "https://ndownloader.figshare.com/files/2607392"], "description"=>"<div><p>Background</p><p>The contribution of individuals with subclinical infection to the transmission and endemicity of cutaneous leishmaniasis (CL) is unknown. Immunological evidence of exposure to <i>Leishmania</i> in residents of endemic areas has been the basis for defining the human population with asymptomatic infection. However, parasitological confirmation of subclinical infection is lacking.</p><p>Methods</p><p>We investigated the presence and viability of <i>Leishmania</i> in blood and non-invasive mucosal tissue samples from individuals with immunological evidence of subclinical infection in endemic areas for CL caused by <i>Leishmania</i> (<i>Viannia</i>) in Colombia. Detection of <i>Leishmania</i> kDNA was conducted by PCR-Southern Blot, and parasite viability was confirmed by amplification of parasite 7SLRNA gene transcripts. A molecular tool for genetic diversity analysis of parasite populations causing persistent subclinical infection based on PCR amplification and sequence analysis of an 82bp region between kDNA conserved blocks 1 and 2 was developed.</p><p>Principal Findings</p><p>Persistent <i>Leishmania</i> infection was demonstrated in 40% (46 of 114) of leishmanin skin test (LST) positive individuals without active disease; parasite viability was established in 59% of these (27 of 46; 24% of total). Parasite burden quantified from circulating blood monocytes, nasal, conjunctival or tonsil mucosal swab samples was comparable, and ranged between 0.2 to 22 parasites per reaction. kDNA sequences were obtained from samples from 2 individuals with asymptomatic infection and from 26 with history of CL, allowing genetic distance analysis that revealed diversity among sequences and clustering within the <i>L</i>. (<i>Viannia</i>) subgenus.</p><p>Conclusions</p><p>Our results provide parasitological confirmation of persistent infection among residents of endemic areas of <i>L</i>. (<i>Viannia</i>) transmission who have experienced asymptomatic infection or recovered from CL, revealing a reservoir of infection that potentially contributes to the endemicity and transmission of disease. kDNA genotyping establishes proof-of-principle of the feasibility of genetic diversity analysis in previously inaccessible and unexplored parasite populations in subclinically infected individuals.</p></div>", "links"=>[], "tags"=>["Cutaneous Leishmaniasis BackgroundThe contribution", "leishmanin skin test", "parasitological confirmation", "parasite viability", "subclinical infection", "pcr", "individual", "Diversity analysis", "lst", "kdna", "mucosal tissue samples", "82 bp region", "developed.Principal FindingsPersistent Leishmania infection", "parasite populations", "cl", "parasite 7 SLRNA gene transcripts"], "article_id"=>1623852, "categories"=>["Biological Sciences", "Ecology"], "users"=>["Mariana Rosales-Chilama", "Rafael E. Gongora", "Liliana Valderrama", "Jimena Jojoa", "Neal Alexander", "Luisa C. Rubiano", "Alexandra Cossio", "Emily R. Adams", "Nancy G. Saravia", "María Adelaida Gómez"], "doi"=>["https://dx.doi.org/10.1371/journal.pntd.0004273.s001", "https://dx.doi.org/10.1371/journal.pntd.0004273.s002", "https://dx.doi.org/10.1371/journal.pntd.0004273.s003", "https://dx.doi.org/10.1371/journal.pntd.0004273.s004", "https://dx.doi.org/10.1371/journal.pntd.0004273.s005", "https://dx.doi.org/10.1371/journal.pntd.0004273.s006"], "stats"=>{"downloads"=>4, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/Parasitological_Confirmation_and_Analysis_of_Leishmania_Diversity_in_Asymptomatic_and_Subclinical_Infection_following_Resolution_of_Cutaneous_Leishmaniasis/1623852", "title"=>"Parasitological Confirmation and Analysis of <i>Leishmania</i> Diversity in Asymptomatic and Subclinical Infection following Resolution of Cutaneous Leishmaniasis", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2015-12-17 08:09:00"}

PMC Usage Stats | Further Information

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Relative Metric

{"start_date"=>"2015-01-01T00:00:00Z", "end_date"=>"2015-12-31T00:00:00Z", "subject_areas"=>[]}
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