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Malaria 2
Clinical Parasitology
FACTM (Clinical) Pt 1
Tim Inglis
Division of Microbiology & Infectious Diseases,
PathWest Laboratory Medicine, WA
Malaria series
Malaria 1 Clinical Tropical Medicine
Malaria 2 Clinical Parasitology
Malaria 3 Public Health, Travel & Expedition Medicine
Malaria 4 Clinical Entomology
Study materials
Practical Guide to Diagnostic Parasitology.
2nd
Edition, Garcia LS. 2009. ASM Press.
• Recommended bookshelf
• FACTM study notes
• Specialist review articles
• Self-assessment questions
Blog http://micrognome.priobe.net
Web www.priobe.net
FACTM http://lifeinthefastlane.com/exams/actm-fellowship/
Parasitology introduction
Plasmodium
– protozoan – unicellular, nucleated Sporozoan, Apicomplexa
– life cycle – multiple stages in mammals or birds & mosquito
– asexual cycle – schizogony in animals
– sexual reproduction – sporogony, requires mosquito
Life cycle
MOSQUITO HUMAN
Investigations
• Key questions:
– Does the patient have malaria?
– Does the patient have P. falciparum malaria?
– Does the patient have another infection?
• Blood films: at least 2, preferably 3 at intervals by 2 methods
(thick & thin ), for parasite detection, count, determination of
species & stages present
• Rapid tests: dipstick for P. falciparum histidine-rich protein II,
quantitative buffy coat, and PCR assays; mainly for P. falciparum
infection, not useful for parasite count
Rapid antigen test
• HRP II:
– only useful for P. falciparum
– may detect P. vivax
– more sensitive than thin film
– comparable with thick film
– be aware of potential FP
– supplements blood film
• Alternative rapid tests:
– QBC
– real time PCR
Blood films
2 methods
– Thick
single drop of blood & Geimsa stain
around 30x as sensitive as thin film
requires experience to interpret
may pick up mixed infection
– Thin
less sensitive but easier to read
examine optimal part of blood film
parasite features easier to recognise
needs repetition x 2-3
% RBC affected
can miss low level parasitaemia and dual infections
Plasmodium recognition chart
KEY
1-4 trophozoites
5 schizont
6 macrogamete
7 microgamete
P.f Plasmodium falciparum
P.v Plasmodium vivax
P.m Plasmodium malariae
P.O Plasmodium ovale
Inglis TJJ. Churchill’s Pocketbook of Clinical Microbiology.
Churchill Livingstone, Edinburgh, 1997.
P. falciparum
• trophozoite: accolé form, multiple per RBC,
double chromatin dots, Maurer’s dots, crenated
RBC, high % RBCs infected
• schizont: rarely seen, except severe
infections, tightly clumped merozoites, malaria
pigment
• gametocytes: crescent or banana-shaped
• dual infection with P. vivax may not show up on
thin film
P. vivax
• trophozoite: single per RBC, single
chromatin dots in early forms, thicker ring
forms, Schuffner’s (pink) dots, amoeboid
cytoplasm, lower % RBCs infected, shape of
RBC unchanged
• schizont: more often seen, less tightly
clumped 12-24 merozoites, malaria pigment
• gametocytes: ovoid shaped, if seen
P. ovale
• trophozoite: thicker ring forms,
oval shaped RBCs, prominent Shuffner’s dots,
• schizont: 4-14 merozoites, RBC has
jagged edge, irregular merozoite arrangement
• gametocyte: irregular nucleus
P. malariae
• trophozoite: thicker ring forms, sometimes
central ‘owl’s eye appearance, stippling is rare,
late band form with prominent malaria pigment
• schizont: 6-12 merozoites in rosette formation
around malaria pigment
• gametocyte: microgametocyte has compact
nucleus
Emerging issues
1. Mechanisms of Plasmodium pathogenesis
2. Where P. falciparum goes
3. Vulnerability points in the life cycle
4. Immunity to Plasmodium spp.
5. Zoonotic malaria: P.knowlesi
Malaria series
Malaria 1 Clinical Tropical Medicine
Malaria 2 Clinical Parasitology
Malaria 3 Public Health, Travel & Expedition Medicine
Malaria 4 Clinical Entomology

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Factm malaria 2

  • 1. Malaria 2 Clinical Parasitology FACTM (Clinical) Pt 1 Tim Inglis Division of Microbiology & Infectious Diseases, PathWest Laboratory Medicine, WA
  • 2. Malaria series Malaria 1 Clinical Tropical Medicine Malaria 2 Clinical Parasitology Malaria 3 Public Health, Travel & Expedition Medicine Malaria 4 Clinical Entomology
  • 3. Study materials Practical Guide to Diagnostic Parasitology. 2nd Edition, Garcia LS. 2009. ASM Press. • Recommended bookshelf • FACTM study notes • Specialist review articles • Self-assessment questions Blog http://micrognome.priobe.net Web www.priobe.net FACTM http://lifeinthefastlane.com/exams/actm-fellowship/
  • 4. Parasitology introduction Plasmodium – protozoan – unicellular, nucleated Sporozoan, Apicomplexa – life cycle – multiple stages in mammals or birds & mosquito – asexual cycle – schizogony in animals – sexual reproduction – sporogony, requires mosquito
  • 6.
  • 7. Investigations • Key questions: – Does the patient have malaria? – Does the patient have P. falciparum malaria? – Does the patient have another infection? • Blood films: at least 2, preferably 3 at intervals by 2 methods (thick & thin ), for parasite detection, count, determination of species & stages present • Rapid tests: dipstick for P. falciparum histidine-rich protein II, quantitative buffy coat, and PCR assays; mainly for P. falciparum infection, not useful for parasite count
  • 8. Rapid antigen test • HRP II: – only useful for P. falciparum – may detect P. vivax – more sensitive than thin film – comparable with thick film – be aware of potential FP – supplements blood film • Alternative rapid tests: – QBC – real time PCR
  • 9. Blood films 2 methods – Thick single drop of blood & Geimsa stain around 30x as sensitive as thin film requires experience to interpret may pick up mixed infection – Thin less sensitive but easier to read examine optimal part of blood film parasite features easier to recognise needs repetition x 2-3 % RBC affected can miss low level parasitaemia and dual infections
  • 10. Plasmodium recognition chart KEY 1-4 trophozoites 5 schizont 6 macrogamete 7 microgamete P.f Plasmodium falciparum P.v Plasmodium vivax P.m Plasmodium malariae P.O Plasmodium ovale Inglis TJJ. Churchill’s Pocketbook of Clinical Microbiology. Churchill Livingstone, Edinburgh, 1997.
  • 11. P. falciparum • trophozoite: accolé form, multiple per RBC, double chromatin dots, Maurer’s dots, crenated RBC, high % RBCs infected • schizont: rarely seen, except severe infections, tightly clumped merozoites, malaria pigment • gametocytes: crescent or banana-shaped • dual infection with P. vivax may not show up on thin film
  • 12. P. vivax • trophozoite: single per RBC, single chromatin dots in early forms, thicker ring forms, Schuffner’s (pink) dots, amoeboid cytoplasm, lower % RBCs infected, shape of RBC unchanged • schizont: more often seen, less tightly clumped 12-24 merozoites, malaria pigment • gametocytes: ovoid shaped, if seen
  • 13. P. ovale • trophozoite: thicker ring forms, oval shaped RBCs, prominent Shuffner’s dots, • schizont: 4-14 merozoites, RBC has jagged edge, irregular merozoite arrangement • gametocyte: irregular nucleus P. malariae • trophozoite: thicker ring forms, sometimes central ‘owl’s eye appearance, stippling is rare, late band form with prominent malaria pigment • schizont: 6-12 merozoites in rosette formation around malaria pigment • gametocyte: microgametocyte has compact nucleus
  • 14. Emerging issues 1. Mechanisms of Plasmodium pathogenesis 2. Where P. falciparum goes 3. Vulnerability points in the life cycle 4. Immunity to Plasmodium spp. 5. Zoonotic malaria: P.knowlesi
  • 15. Malaria series Malaria 1 Clinical Tropical Medicine Malaria 2 Clinical Parasitology Malaria 3 Public Health, Travel & Expedition Medicine Malaria 4 Clinical Entomology