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MALARIAL SMEAR, COUNTING AND REPORTING

QuestionAnswer
Malaria parasites - FVOM Plasmodium falciparum Plasmodium vivaxPlasmodium ovalePlasmodium malariae
Developmental stages seen in Blood films: 1. Trophozoite2. Schizont 3. Gametocyte
RED CELL MORPHOLOGY 1. Size of infected RBCs 2. Infection of RBCs3. Presence of stipplings/dots
- Maurer’s dot- Coarse in few in the cytoplasm P. falciparum
- Schuffner’s dot- Fine & abundant P. vivax
- James dot- Coarse & abundant P. ovale
- Ziemann’s dot- Few and fine P. malariae
Size of infected RBCs  Not enlarged: P. falciparum & P. malariae
Size of infected RBCs  Large: P. vivax & P. ovale
Infection of RBCs  No. of trophozoites- Single: P. vivax
Infection of RBCs  No. of trophozoites- Multiple: P. falciparum
LABORATORY TECHNIQUES IN MALARIA MICROSCOPY  THICK SMEAR THIN SMEAR
- Layer of red blood cells 10-20 times thicker than a thin film - Parasites are within red cell ghosts (staining unfixed red cells causes hemolysis) - Used to detect parasites and estimates parasite density - Gives sensitivity to diagnosis THICK SMEAR
THICK SMEAR - Layer of red blood cells ? times thicker than a thin film 10-20
- Single layer of red blood cells - Used as label to identify patient- Used to identify parasites species, after they have been seen in thick film - Gives specificity to diagnosis THIN SMEAR
a lead pencil to write the following information on the frosted end of the slide: patient’s name, age, sex, laboratory code and date of collection
Clean the ? or 4th finger from the thumb with cotton soaked in ?, using firm strokes to remove dirt and grease from the finger. Air dry the finger 3rd , 4th.. 70% alcohol
Using a sterile lancet, puncture the ball of the patient’s finger. Apply gentle pressure to the finger to express the ? drop of blood and wipe it away with dry cotton first ,
Apply gently pressure to the finger then collect ? small drops (approximately 6 uL) of blood on one side of a clean slide 3
Collect? small drop (appx. 2 uL) next to the 3 drops, leaving some space between the thick and thin smears to be made. Handle clean slides only by the edges 1
Make a thin smear first. Place slide on a flat, firm stage. Bring down a second slide (spreader) on the 1 small drop at an angle of ? degree, allow the blood to run along its edge 45
Fix the thin smear with ? by using a proper or by pipping it in the solution for a few seconds methanol
Air dry the smear. Do not fix thick smear TRUE
Stain thick and thin smears with ? for ? mins (10% Giemsa stain preparation: 1 part of stain + 9 parts of buffered water) Giemsa stain ,, 8-10
10% (Rapid method) - Individual flooding - Giemsa stock 9 drops
10% (Rapid method) - Individual flooding - Buffered H2O 3 mL
10% (Rapid method) - Individual flooding - Staining time 8-10 mins
10% (Rapid method) - Mass (immersion/coplin jar) - For 50 ml - Giemsa stock 5 ml
10% (Rapid method) - Mass (immersion/coplin jar) - For 50 ml - Buffered H2O 45 ml
10% (Rapid method) - Mass (immersion/coplin jar) - For 50 ml - Staining time 8-10 mins
Place the slide in drying rack in with the ? DOWNto drain and dry thick film
Position the thick film in line with the ? objective lens and adjust the light optimally 10x
Place a drop of ? on the thick film,scan and select a well-stained, even portion of the blood film immersion oil
Parasite count can be estimated from either:  THICK FILM No. of parasites/ uL of blood
Parasite count can be estimated from either:  THIN FILM No. of parasites/ cu. mm blood
2 TYPES OF METHOD: 1. CROSS SECTIONAL METHOD (recommended technique in parasite counting) 2. BATTLEMENT METHOD
FORMULA: No. of parasites x 8,000200 WBCs = no. of parasites/ uL of blood
separate the counts of Pv (any/all stages seen) from sexual forms (gametocytes) of P. falciparum Vfg
count the parasites of Pv and Pf as one Vf –

Created by: blues1234

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