Download Introduc8on` Materials`and`methods` Conclusions` Results

Rojelio'Mejia1,'Ashish'Damania1,'
•  !>2"billion"GI"parasite"infec3ons"worldwide""
•  Poorest"and"resource8deprived"communi3es""
•  Standard"method"of"diagnosis:"Stool"microscopy"
•  Sensi3vity"variable"depending"on"prevalence,"
species,"and"concentra3on"method"
•  50890%"sensi3vity""
•  Underes3mates"polyparasi3sm"
•  qPCR"is"rapid,"quan3ta3ve,"and"high8throughput""
species8specific"method"
•  GI"parasites"may"disrupt"normal"intes3nal"microbiota"
•  Decreased"biodiversity"is"associated"with"disease"
•  Malabsorp3on"
•  Inflammatory"bowel"diseases""
Results""
•  qPCR"iden3fies"more"polyparasi3sm"than"
microscopy"
Microscopy"""""""""""""""""""""""""""""""""""""""""""""""""qPCR"
qPCR"(ITS"region)"(ABI"7500)"iden3fied"more"cases"
of"Ascaris,"hookworm"(Hw),"Strongyloides,'
Entamoeba'histoly=ca"and"Giardia"infec3on"than"
microscopy.""""(Tt,"C)"no"posi3ves"
• 
80
*'
Prevalence (%)
Introduc8on'
,
60
0
qPCR %
Microscopy %
• 
*'
•  Giardia"infected"group"had"higher"abundance"
of"Bacteroidetes"compared"to"No"Parasites"
group"with"higher"Firmicutes'(p"<"0.05)'
*'
20
*'
Al
Ad
Na
Hw
Ss
Eh
•  Temperate"climate""
•  99"pa3ent"samples"
Gl
G
None
qPCR"can"detect"more"parasites"than"microscopy"
• 
Ascaris"91.3%"Sens,"90.5%"NPV"
• 
Hookworm"95.5%"Sens,"98.4%"NPV"
• 
Strongyloides"100%"Sens,"NPV"
• 
Giardia"87.5%"Sens,"97.2%"NPV"
qPCR"can"iden3fy"polyparasi3sm"beger"than"
microscopy"
• 
Important"for"treatment"selec3on""
GI"parasi3c"infec3ons"at"high"prevalence""
• 
qPCR'detected"Giardia"6"x"more"than"
microscopy"
Giardia"infected"group"had"decreased"intes3nal"
microbiota"biodiversity"(p"="0.01667)"
• 
Giardia"infected"group"(2.7)"
• 
No"Parasite"group"(3.45)"
Giardia'infected'group"had"significant"increases"in"
Bacteroidetes"specifically"Prevotella'species"
Useful"for"epidemiology"and"morbidity"studies"
• 
Surveillance"ajer"mass"drug"administra3on"
and"vaccine"programs"
• 
Expand"understanding"of"morbidity"and"
malnutri3on"
• 
Cost"is"less"than"$1.00"US"per"pa3ent"to"
screen"for"these"parasites"
Future"direc3ons"
• 
Correlate"quan3ty"of"parasite"DNA"with"
clinical"outcomes"
• 
Associate"morbidity"to"changes"in"
microbiome"
• 
Treat"children"with"an38parasi3cs"and"
evaluate"changes"in"microbiome"
P
2.5e+07
R3
• 
2.0e+07
Actinobacteria
•  f"
•  Decreased"intes3nal"bacterial"biodiversity"in"combined'
Giardia"group"versus"No"parasite"group"
Abundance
Bacilli
Bacteroidetes
1.5e+07
Betaproteobacteria
• 
Clostridia
delta/epsilon subdivisions
1.0e+07
Erysipelotrichia
Gammaproteobacteria
Negativicutes
5.0e+06
Spirochaetia
Verrucomicrobia
0.0e+00
No"Parasite"group"
Shannon Alpha Diversity
Giardia"group"
4.0
p = 0.01667
3.5
Spirochaetes
•  Giardia"infec3on"decreases"intes3nal"
bacterial"biodiversity"
Proteobacteria
R2
Firmicutes
• 
Bacteroidetes/Chlorobi group
Actinobacteria
Spirochaetes
Proteobacteria
Firmicutes
Chlamydiae/Verrucomicrobia group
Bacteroidetes/Chlorobi group
Actinobacteria
Spirochaetes
Proteobacteria
Firmicutes
Chlamydiae/Verrucomicrobia group
Bacteroidetes/Chlorobi group
Actinobacteria
•  No"recent"an3bio3cs"
Chlamydiae/Verrucomicrobia group
•  Asymptoma3c"children"
•  Ages"2810"years"old"
•  Stool"samples"evaluated"by"qPCR"and"microscopy"for"
presence"of":""
Ascaris'lumbricoides"(Al)''''''''''Strongyloides'stercoralis'(Ss)"
Ancylostoma'duodenale"(Ad)""Giardia'lamblia"(Gl)"
Necator'americanus'(Na)"""""""""Cryptosporidium"species"(C)"
Trichuris'trichiura'(Tt)'''''''''''''''Entamoeba'histoly=ca'(Eh)"
•  qPCR"required"50"mg"samples"
•  Microscopy"required"2"g"samples"
•  McMaster"technique"(semi8quan3ta3ve)""
•  NEBNext®"Microbiome"DNA"Enrichment"Kit"
•  NEBNext®"Ultra™"DNA"Library"Prep"Kit"for"Illumina®"
•  Illumina"MiSeq®"“shotgun”"sequencing"
• 
Giardia'''''''''''''''''''''''No"Parasites'
Materials'and'methods'
•  Field"site:"Orán,"Argen3na""
•  Peri8urban"community""
• 
• 
*P"<"0.05"
40
Conclusions'
3.0
Acknowledgements'
2.5
2.0
Giardia infected
No Parasites
Funding"for"this"project"was"provided"by"the"Na3onal"
School"of"Tropical"Medicine"and"New"England"BioLabs,"Inc."
!