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To establish the diagnosis of malaria, a blood film must be prepared from fresh blood obtained by pricking the
patient's finger (Figures A-1 and A-2).* The thin film is fixed in methanol before staining; the thick film is stained unfixed.
Certain hospitals have a Wright-Giemsa stain available, which is acceptable; however, Wright stain alone will not reliably
indicate Plasmodium parasites. For best results, the film should be stained with a 3% Giemsa solution (pH of 7.2) for 30--45
minutes. In P. falciparum infections, the parasite density should be estimated by counting the percentage of red blood cells infected
--- not the number of parasites --- under an oil immersion lens on a thin film.
Thick blood films are more sensitive in detecting malaria parasites because the blood is concentrated, allowing a
greater volume of blood to be examined. However, thick films are more difficult to read, and thin films might be preferred
by laboratories that have limited experience.
Plasmodium parasites are always intracellular, and they demonstrate, if
stained correctly, blue cytoplasm with a red chromatin dot. Common errors in reading malaria films are caused by platelets
overlying a red blood cell, concern regarding missing a positive slide, and misreading artifacts as parasites. Persons suspected of
having malaria, but whose blood films do not indicate the presence of parasites, should have blood films repeated
approximately every 12--24 hours for 3 consecutive days. If films remain negative, then the diagnosis of malaria is unlikely.
For rapid diagnosis, the thick and thin films should be made on separate slides. The thin film should be air-dried,
fixed with methyl alcohol, and immediately stained. If no parasites are visible on the thin film, the laboratorian should wait
until the thick film is dry, then examine it for organisms that might not have been detected on the thin preparation.
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