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Laboratory Diagnosis
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of Human Parasitic Infections
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Eric Brienen
Annelies van Goor
Lisette van Lieshout
& guest lectures
[email protected]
ar
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u
Department of Parasitology
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Leiden University
Centre
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During the course: +31 (0)6- 1301 5374
Department of Parasitology
Leiden University Medical Centre
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Department of Parasitology
Leiden University Medical Centre
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Practical aspects ra
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9:00 – 17:00
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Coffee/tea; lunch
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Hand-outs presentations, ESCMID online
of week1, book
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Small adaptations of program
possible
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a
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y of the day & participants
O patient
Real-life cases (blinded);
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can bring in
2)
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Microscopy
– arrangements/duo’s; ENG/NL
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Photo
E quizzes
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Department of Parasitology
Leiden University Medical Centre
Common intestinal worms
Schistosomiasis
Laboratory procedures faeces
Zoönotic helminths
•
•
•
•
Common intestinal protozoa
Artefacts
Malaria
Laboratory procedures blood
LU
M
New developments
Coccidian & rare parasites
Blinded & Clinical cases
(E-learning)
Department of Parasitology
Leiden University Medical Centre
Ecto-parasites
Delusions of parasitosis
Parasites and internet
Research (LUMC visit)
Opportunistic infections
-P
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•
•
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Skin parasites
More malaria, alternative diagnosis
More clinical aspects
More treatment
ar
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Week 1
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Layout program
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Week 2
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Pneumocystosis; toxoplasmosis
Filariasis, tissue nematodes
Trypanosomiasis
Leishmaniasis
Free-living amebic-infections
LU
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as
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gy
What is a parasite? ary
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gy
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as
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LU
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Large parasites?? ry
a
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b
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Longest?
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Human tape worms - Cestoda
Diphyllobothrium latum
C
LU
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as
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to
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gy
Large parasites?? ary
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http://images-02.delcampe-static.net/img_large/auction/000/078/231/652_001.jpg
>10 meter
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LU
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Dracunculus medinensis
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guinea worm disease; female worm 70-120 cm
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http://www.theatlantic.com/health/archive/2015/01/carter-center-guinea-worm-jimmy-carter-parasite-pipe-filter/384557/
LU
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Small parasite?? ry
a
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b
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Single cell parasites
L
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(Protozoa)
u
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Intra/extra cellular in
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Intestinal ID
©
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Blood
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Tissue
C
LU
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What is a parasite? ary
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“ An organism that lives
u
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c
in or on and takes its
e r
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nourishment from e
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another organism.
a
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A parasite D
cannot live
I
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independently.”
M
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LU
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What is a parasite? ry
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b
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From the Latin parasitus,
L
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r
the latinisation
of
u
t
c
the Greek παράσιτος
(parasitos),
e
r
L
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"one who eatsnat the table
of another"
t
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I
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at
the expense of the other, the host”
C
S
feeding, protection, replication etc.
LU
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Cell Structure parasiteary
r
b
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Human parasitology:
e
r
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t
Eukaryote
c
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L
• Cell membrane
o
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• Nucleus
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• CytoplasmaO by
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• Cell organelles
M
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Single cell
Multiple cellular
gy
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LU
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Most common parasite??
r
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In the Netherlands - number of new cases/year?
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Mother hunting for headlice, detail of a
painting by Jan Siberechts
A heated air device
designed to kill headlice
via desiccation.
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Most dangerous parasite??
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World wide - number of casualties /year? L
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WHO : In 2010, malaria caused an estimated 660 000 deaths (with an uncertainty
range of 490 000 to 836 000), mostly among African children.
gy
y
Most dangerous parasite??
r
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Transmission cycle
L
Plasmodium
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≈1870 parasite in blood
Air, inhalation?
Water, ingestion?
Human: intermediate host
LU
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1897 - Ronald Ross
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Mosquito: definitive host
LU
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Parasites depend on host(s)
r
a
r
Host = (in)vertebrate
species
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L
e1
parasite
Host
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parasite
Host 1
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Host 3
C
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parasite
Host 1
Host 2
Human - Host
zoönotic infection (normally in other mammalians)
LU
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Complexity of life cycles
r
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L
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I
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C
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Plasmodium
Schistosoma
LU
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to
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Complexity of life cycles
r
a
r
b
i
L
e
r
u
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c
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L
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Knowledge
essential
for:
e
h
t
n
i
u
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•
distribution
a
n
y
O • treatment
b
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• prevention
M
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Plasmodium
Schistosoma
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After malaria……. ary
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LU
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http://blogs.plos.org/speakingofmedicine/2015/01/16/one-million-deaths-parasites/
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Range of parasitic infections
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Classification of parasites
r
a
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L
Endo-parasite
Ecto-parasite
e
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L
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Scabies mite
Entamoeba histolytica
Schistosoma
LU
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ar
Protozoa
Helminths
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Characteristics of: ary
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b
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Protozoa:L
e
r
u
t
c
•
single cell organism
e
r
L
• intra/extra
cellular
o
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h
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•
1-100
µm
n
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• multiply within host
I
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• binary fission ….
Giardia
• …or schizogony/sporogony
LU
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Characteristics of helminths
r
a
r
Adult worm: Lib
• mm to meters
e
r
• multi
cellular
u
t
c organs
•e
multiple
r
L
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e
h
t
•
stable
population in single host
n
i
u
l
•amostly chronic
n
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I
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• egg or larvae production
C
• eggs/larvae leave the host to
S
Schistosoma
establish a new infection/worms
LU
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Parasites = tropical infection?
r
a
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Parasites in the Netherlands?
r
a
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b
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L
e
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L
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S
E
ar
as
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to
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gy
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Microscopy “gold” standard
r
a
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e
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(+)......
I
(+).......
M
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(-).......
E
LU
M
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Van Leeuwenhoek, 1681
(-)........
LU
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Diagnostic sampling? ry
a
r
b
i
L
e
r
u
t
c
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L
o
e
h
t
n
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a
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I
M
C
S
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gy
y
Microscopy procedures
r
a
r
b
i
L
e
r
u
t
c
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Demonstration during labpractical:
L
o
1a - Direct smear / Ridley
e
h
t
1b - Baermann
/
Coproculture
n
i
u
l
a
n
2 - KK/urine
y
O filtration/Glycerin
b
D
I
©
3 - Iodine/Eosine
M
C
S 4 - Blood
LU
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as
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to
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Fixative & permanent staining
5 – ZN (& others?)
C
LU
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Alternatives? ry
a
r
b
i
L
I think it is
e
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a
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mammothec
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L
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Let's first do
a PCR to
confirm your
findings
gy
Diagnosis of
y
r
a
Parasitic Infectious Diseases
r
b
to
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I
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O
ar
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Microscopy
r
u
t
Serology (Ab detection)ec
r
L
o
Antigen detectionne
h
t
i
u
l
a
n (q-PCR)
Molecular diagnosis
by
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1.
2.
3.
4.
i
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e
©
LU
M
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• IdealCdiagnostic assay
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• Parasite
&......
• => target population
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ar
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to
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Diagnostic aims ary
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LU
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Patient or “infected individual”
LU
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Ideal diagnostic assay: ry
a
r
b
i
 Sensitive and specific
L
e uniform
 Test results: reproducible, simple,
r
u
t
(quantitative?)
c
e r
L
o
e
h
 Capable to process
large sample
numbers
t
n
i
u
l
a
n
 Fast
y
O
b
 Easy to perform
(labour?)
D
I
©
 FieldM
applicable
C
S
E Affordable (costs, labour)
 Available
LU
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a
Treatment of parasitic infections
r
b
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S
Department of Parasitology
Leiden University Medical Centre
LU
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C
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ar
as
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to
lo
gy
So parasites are....... ry
a
r
b
i
 Eukaryote organism
L
 (Partly) dependent on host(s) re
u
t
c
 Transmission cycle – adaptations/stages
e r
L
 Mostly diagnosed by e
microscopy
o
h
t
n
i
u
 More common in lthe
tropics
a
n
y
 Common (?) O
b
≈ 150 speciesID
described©
in humans
M
≈ a.o. 200
million malaria infections
C
≈ a.o.
2 billion infected with soil-transmitted helminths
S
E≈ 3 (/7) billion infected with chronic parasite infection
 Burden of disease?
LU
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to
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gy
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At the end of the course…
r
a
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