3/27/23, 10:11 PM CDC - DPDx - Diagnostic Procedures - Stool Specimens

DPDx - Laboratory Identification of Parasites of Public Health Concern

DPDx - Laboratory Identification of Parasites of Public Health

Concern

Stool Specimens - Specimen Collection

1. Collect the stool in a dry, clean, leakproof container. Make
sure no urine, water, soil or other material gets in the
container.
2. The image on the right demonstrates the distribution of
protozoa in relation to stool consistency and should be taken
into consideration when specimens are received.
3. Fresh stool should be examined, processed, or preserved
immediately. An exception is specimens kept under
refrigeration when preservatives are not available; these
specimens are suitable for antigen testing only.
Distribution of protozoa in relation to stool consistency
4. Preserve the specimen as soon as possible. If using a
commercial collection kit, follow the kit’s instructions. If kits
are not available, the specimen should be divided and stored in two different preservatives, 10% formalin and PVA
(polyvinyl-alcohol), using suitable containers. Add one volume of the stool specimen to three volumes of the
preservative.
5. Insure that the specimen is mixed well with the preservative. Formed stool needs to be well broken up.
6. Insure that the specimen containers are sealed well. Reinforce with parafilm or other suitable material. Insert the
container in a plastic bag.
7. Certain drugs and compounds will render the stool specimens unsatisfactory for examination. The specimens should be
collected before these substances are administered, or collection must be delayed until after the effects have passed.
Such substances include: antacids, kaolin, mineral oil and other oily materials, non-absorbable antidiarrheal
preparations, barium or bismuth (7-10 days needed for clearance of effects), antimicrobial agents (2-3 weeks), and
gallbladder dyes (3 weeks).
8. Specimen collection may need to be repeated if the first examination is negative. If possible, three specimens passed at
intervals of 2-3 days should be examined.

Preservation of specimens is necessary when stool specimens cannot be examined within the prescribed time interval.
Various preservatives are available (see table), with the two most commonly used being 10% aqueous formalin and PVA
(polyvinyl-alcohol). If molecular detection (PCR) is required, refer to the molecular diagnosis section to obtain specific
information on how to collect, preserve, and ship the specimens.

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3/27/23, 10:11 PM CDC - DPDx - Diagnostic Procedures - Stool Specimens

Preservative Advantages Disadvantages

10% Formalin All purpose fixative Not suitable for some

Easy to prepare permanent smears stained with
trichrome
Long shelf life
Inadequate preservation of
Good preservation of morphology of
morphology of protozoan
helminth eggs, larvae, protozoan cysts, and
trophozoites
coccidia
Can interfere with PCR,
Suitable for concentration procedures and
especially after extended
UV fluorescence microscopy
fixation time
Suitable for acid-fast, safranin, and
chromotrope stains
Compatible with immunoassay kits and UV
fluorescence microscopy

MIF Components both fix and stain organisms Not suitable for some
merthiolate-iodine- Easy to prepare permanent smears stained with
formaldehyde) trichrome
Long shelf life
Inadequate preservation of
Useful for field surveys
morphology of protozoan
Suitable for concentration procedures trophozoites
Iodine interferes with other
stains and fluorescence
Iodine may cause distortion of
protozoa

LV-PVA Good preservation of morphology of Inadequate preservation of

(low viscosity polyvinyl- protozoan trophozoites and cysts morphology of helminth eggs
alcohol) Easy preparation of permanent smears and larvae, coccidia, and
stained with such as trichrome (solution microsporidia
both preserves organisms and makes them Contains mercuric chloride
adhere to slides) Difficult and expensive to
Preserved samples remain stable for dispose of
several months Difficult to prepare in the
laboratory
Not suitable for concentration
procedures
Cannot be used with
immunoassay kits
Not suitable for acid-fast,
safranin and chromotrope
stains

SAF Suitable for both concentration procedures Requires additive (e.g., albumin-
(sodium acetate-acetic acid- and preparation of permanent stained glycerin) for adhesion of
formalin) smears specimens to slides
Easy to prepare Permanent stains not as good
Long shelf life as with PVA or Schaudinn’s
fixative
Suitable for acid-fast, safranin, and
chromotrope stains
Compatible with immunoassay kits

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3/27/23, 10:11 PM CDC - DPDx - Diagnostic Procedures - Stool Specimens

Preservative Advantages Disadvantages

Schaudinn’s Fixative Good preservation of morphology of Less suitable for concentration

protozoan trophozoites and cysts procedures
Easy preparation of permanent stained Contains mercuric chloride
smears Inadequate preservation of
morphology of helminth eggs
and larvae, coccidia, and
microsporidia
Poor adhesion of liquid or
mucoid specimens to slides

Modified PVA Permanent smears can be made and Staining not consistent
copper or zinc stained with trichrome Organism morphology may be
Zinc is preferred over copper poor
No mercuric chloride Copper-morphology of cysts and
trophozoites is poor
Zinc-better morphology but not
comparable to LV-PVA

One-Vial Fixatives Concentrate and permanent smear can be Certain one-vial fixatives must
(such as Ecofix, Parasafe, made out of one vial use certain stains
Unifix, Proto-fix, STF, and Immunoassays can be done on most Color difference of stain
others that may be available)
No mercuric chloride Staining not always consistent
Sometimes more expensive
than formalin and LV-PVA

Because 10% formalin and PVA have complementary advantages (see table), it is recommended that the specimen be divided
and preserved in both types of preservatives (add one volume of stool to three volumes of the preservative). Commercial
two-vials kits are available for this purpose. Preserved specimens can be stored for several months.

For additional information on stool collection, call the Division of Parasitic Diseases at (404) 718-4110.
Page last reviewed: May 3, 2016

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