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Specimen Collection

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Parasitology Specimen Collection Guidelines

Specimen Type	Collection^a					Transport Time and Temp to lab
Specimen Type	Guidelines	Device	Preservative	Minimum Vol	Replica limits	Transport Time and Temp to lab	Comments
Arthropod (Tick) for ID	Place in leakproof container with 70% alcohol	Leakproof container	70 % alcohol	Entire Arthropod	None	Indefinite at RT	See separate collection procedure for scabies.
Blood: Direct smear	1. Warm the patient's hands by covering them with a hot moist towel, by immersing them in warm water, or by rubbing them together briskly. 2. Disinfect the palmar surface of the tip of the middle or "ring" finger with gauze soaked with 70% alcohol (do not use cotton as it may introduce artifacts). 3. Allow alcohol to dry completely, as residual alcohol does not permit a drop of blood to "round up" and may also fix the Gold cells, rendering the thick smear unsuitable for staining. 4. Puncture the palmar area with a sterile disposable lancet, resulting in free flowing blood.	Wear gloves when preparing thin or thick films. Thin-smear preparation: 1. Place one drop of blood near one end of a slide. 2. Hold another slide at 45^o angle and draw it into the drop of blood. 3. Allow the blood to spread the width of the slide and then rapidly push the spreader slide to the opposite end, producing a feathered smear. 4. Label slide, dry at RT, and stain as soon as visibly dry. Thick-smear preparation: 1. Touch a slide to a drop of blood (rounded up on the finger). 2. Rotate the slide to form a circular film about the size of a nickel. (For blood without anticoagulant, stir blood 20-30 s to prevent formation of a fibrin clot.)				Malaria: STAT Other: < 2 h, RT	Optimal time to obtain smear Babesia spp: any time Brugia malayi ^b: ~midnight Leishmania donovaniv ^b: any time Loa loa^b: ~noon Mansonella ozzardi^b: day or night Mansonella perstans^b: night better than day Plasmodium spp.^c:: between chills Trypanosoma cruzi^b: acute stage Trypanosoma brucei gambiense^b^,d: acute stage Trypanosoma brucei rhodesiense^b^,d: acute stage Wuchereria bancrofti^b: ~midnight^c; additional smears obtained 6, 12, or 24 h after admission may be necessary.
Venipuncture	1. For buffy coat concentration if filariasis, trypanosomiasis, and to a lesser extent leishmaniasis, collect 10 ml. of whole blood with heparin or EDTA (0.002 g/10 ml of blood). 2. Submit directly (< 15 min) to the laboratory at RT. 3. Thick or thin smears should be obtained via finger puncture; see above.	Vacutainer	Heparin: filariasis, Trypanosoma spp. EDTA: malaria; see comments	> 10 ml.	1 day	< 30 min, RT	Common parasites: L.donovani, Trypanosoma spp., microfilariae Venipuncture for malaria is common but not recommended because smears must be made within 1 h to detect stippling. However, this approach is common, and personnel learn to identify with or without stippling.
CSF,CNS	See "Specimen Type: CSF" (Table 1) for specific guidelines for obtaining a CSF specimen.	Sterile tube	None	> 1 ml	None	< 30 min, RT	Common parasites: Acanthamoeba spp, Balamuthia mandrillaris, Echinococcus spp., larval cestodes, microsporidia, Naegleria fowleri, Taenia solium, Toxoplasma gondii, Trypansoma spp.

Specimen Type	Collection^a					Transport Time and Temp to lab
Specimen Type	Guidelines	Device	Preservative	Minimum Vol	Replica limits	Transport Time and Temp to lab	Comments
Duodenal aspirate	Place aspirate in a sterile centrifuge tube and transport it directly (< 15 min) to the laboratory since specimens must be examined within 1 h of collection.	Sterile centrifuge tube	None	> 2 ml	None	< 15 min, RT	Common parasites: Clonorchis sinensis (eggs), Cryptosporidium parvum (oocyst), Giardia lamblia (trophozoite), Isospora belli (oocyst), Strongyloides spp. (larvae)
Eye: corneal scraping for Acanthamoeba spp.	1. Instill two drops of local anesthetic into the conjunctival sac and/or onto the corneal epithelium. 2. Using a sterile spatula, scrape ulcers or lesions and either inoculate a non-nutrient agar plate directly or place scraping in PBS and transport to the laboratory. 3. Apply remaining material to two clean glass slides for staining and fix immediately with 95% ethanol (cysts may become airborne when allowed to air-dry).	Direct inoculation of non-nutrient agar coated with bacterial overlay of PBS.	PBS	None	None	< 24 h, RT	Common parasites: Acanthamoeba spp., Naegleria spp. Also contact lenses, lens cases, and all opened solutions.
Feces: Preserved	1. Pass directly into a clean, dry container. 2. Specimens that cannot be examined within the recommended time must be transferred to an appropriate preservative (SAF). Carefully remove the cap and attached spoon to pick several spoonfuls of the stool, especially from areas that are slimy, bloody, or watery. Place the stool into the vial to the fill line. Mix well and allow to stand at RT for 30 min for adequate fixation. 3. For unpreserved specimens, the transport times must correspond to the recommendations provided below. 4. Submit three specimens over 7-10 days because shedding may be intermittent.	Sterile, leakproof, wide-mouth container.	SAF one vial system. Hold at RT for 30 min for fixation.	1 part feces to 3 parts fixative	1/day	Indefinite, RT	Common parasites: helminths, protozoa Unacceptable stool specimen: (1) Contaminated with urine or water (e.g., from diapers); (2) nonpuncturable or dried specimen; (3) specimens containing bismuth, barium, magnesia, mineral oil, gallbladder dye. The general waiting period necessary to allow substances to clear is 7 days, except for gallbladder dye which may require 21 days.