Malaria Species - pametnorcal.org · Malaria Species • Five Plasmodia species –P. falciparum...
Transcript of Malaria Species - pametnorcal.org · Malaria Species • Five Plasmodia species –P. falciparum...
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Malaria Species • Five Plasmodia species
– P. falciparum
– P. vivax
– P. ovale
– P. malariae
– P. knowlesi
• Majority – P. falciparum (most malaria-associated deaths)
• Tropical Africa, Southeast Asia, Oceania, Haiti, Amazon basin of South America, and Dominican Republic
– P. vivax • Central America, Middle East, and India
– P. knowlesi • South East Asia
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Plasmodium life cycle
www.uptodate.com
Sexual cycle in midgut
of female mosquito
Asymptomatic
Symptomatic
(6-16 days)
P. vivax, P. ovale, P. falciparum
- over 48 hr
P. Malariae
- over 72 hr
P. Knowlesi
- 24 hr
Hypnozoites
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Malaria life cycle
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Diagnostic Tests for Blood Parasites ❑ Blood films (thick and thin); stain with Giemsa.
• Prepare films as quickly as possible after collection from patient (be careful,
some blood parasites are extremely contagious, such as Trypanosomes)
• Heparin leads to morphologic distortion; use EDTA
• Draw blood at 12 h intervals for 36 hours if initial smears negative
• Stain with Giemsa or Wright’s/Giemsa
• Examine thin films all around edges for microfilariae
❑ Immunochromatographic lateral flow EIA for malaria
❑ PCR for malaria, Babesia
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Thick and thin smears
Fix with methanol
Do not fix thick smear
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DD
D
DD
D
DD
D
DD
D
DD
D
Sequestered
P.
f
P.
v
P.
o
Identification of Plasmodium species ring form
trophozoite schizont
gametocyte
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Plasmodium malariae/knowlesi
P. malariae band form trophozoite
P. malariae compact old trophozoite
P. malariae immature schizont
P. malariae young immature schizont
Normal sized RBC
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Morphology of
Malarial Parasites
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Differentiate knowlesii
vs malariae by
molecular methods
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David Bell, Chansuda Wongsrichanalai and John W. Barnwell. Ensuring quality and access for malaria diagnosis: how can it be achieved? Nature Reviews Microbiology. 2006:4;682-695.
Lateral flow immunochromatographic assay
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Limitations of Binax
Persistence of HRP-2 after parasite clearance
Humar et al. detected circulating HRP-2 antigen in
68% of treated patients on day 7, and in 27% it was
still present on day 28 (Am J Trop Med Hyg 1997 56:44)
Gene deletion for the production of HRP-2
False-positive results
Cross-reaction with serum rheumatoid factor, causing
Moody Clin Microbiol Rev. 2002 15:66-78
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DNA extraction Real-time PCR
Plasmodium species identification by real-time PCR
Qiagen blood mini kit
•Qiagen RotorGene •FastStart SYBR Green Master mix •Singleplex reactions
EDTA Blood
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Samples tested (49) n Correctly
identified
Plasmodium genus 52 51 (98.1%)
Plasmodium falciparum 22 21 (95.5%)
Plasmodium ovale 8 8 (100%)
Plasmodium vivax 19 19 (100%)
Plasmodium knowlesi 1 1 (100%)
Plasmodium malariae 6 6 (100%)
•Positive: 56 archived pt samples (SHC and Karolinska Institute)
•Negative: 20 EDTA pt samples from Hematology Lab (4 +ve
1. Pf neg, pan pos
(Seq:Pf)
2. Pf by microscopy,
mixed Pf/Po by
PCR/Seq
1 pan neg and Pf pos, pt
undergoing treatment
(Pf primers more sens)
Accuracy of Plasmodium RT-PCR Reference Standard: Microscopy (n=56)
Lefterova et al JCM 2015 PMID:25972416
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Babesia
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Blood and Tissue Parasites ❑ Blood
▪ RBCs – Plasmodium; Babesia
▪ WBCs – Leishmania; Toxoplasma
▪ Plasma – Microfilaria; Trypanosoma
❑ Bone marrow – Leishmania; Trypanosoma cruzi;
Plasmodium
❑ CNS – Taenia; Echinococcus; Naegleria;
Acanthomoeba; Balamuthia; Toxoplasma;
Trypanosoma; microsporidia
❑ Eyes – Acanthamoeba; Toxoplasma; Loa loa;
microsporidia
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Blood and Tissue Parasites
❑ Lungs – Paragonimus; Cryptosporidium; Echinococcus;
Toxoplasma; Helminth larvae (Ex: Strongyloides)
❑ Muscle – Taenia solium; Trichinella; Onchocerca; T. cruzi;
microsporidia
❑ Skin and skin ulcers – Leishmania; Acanthamoeba;
Onchocerca; microsporidia; Dracunculus
❑ Urogenital – Trichomonas; Schistosoma haemotobium;
microfilaria; microsporidia
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Final Quiz – Which parasite is this?
http://www.cdc.gov/malaria/