Enterobius vermicularis

Cellophane (scotch) tape method

The clear-cellulose tape preparation is the most widely used procedure for the detection of human pinworm
infections.
Adult Enterobius vermicularis worms inhabit the large intestine and rectum; however, the eggs are not normally
found in fecal material. During period of extraintestinal migration, the adult female migrates out the anal opening
and deposits the eggs on the perianal skin, usually during the night.

The eggs, and occasionally the adult female worms stick to the glued (sticky) surface of the cellulose tape. These
cellulose tape preparations are submitted to the laboratory, where they are examined under microscope. Commercial
collection systems pinworm paddles are also available

Time of specimen collection

The specimen is collected from the perianal skin. An early-morning sample, before the patient has bathed, or used
the toilet, is optimal. Up to six successive day morning samples should be collected before a negative result is
issued.
Procedure
1.
Place a strip of clear cellulose tape (adhesive side down) on a microscope slide as follows: Starting ca. 1.5
cm from one end, run the tape toward the same end, and wrap the tape around the slide to the opposite end. Tear
the tape even with end of the slide. Attach a label to the tape at the end torn flush with the slide.
2.
To obtain a sample from the perianal area, peel back the tape by gripping the labeled end, and, with the tape
looped (adhesive side outward) over a wooden tongue depressor that is held firmly against the slide and
extended about 2.5 cm beyond it, press the tape firmly several times against the right and left perianal folds.
3.
Smooth the tape back on the slide, adhesive side down.
4.
Label with patient name and date.
5.
Submit the tapes and slides to the laboratory in a plastic bag.
6.
Once the sample is obtained, remove the slide from the tongue depressor and affix the tape sticky side
down on the glass slide.
7.
Label the slide with the patient name and date of collection
8.
Examine the slide under a microscope using the low power (10x) objective. The eggs can be made more
visible by detaching the tape from the slide, adding a drop of xylene or toluene, and again affixing the tape.
Results
1.
The eggs of E.vermicularis measure 50-60 micrometer x 20-30 micrometer and have a relatively thick,
smooth shell and an ovoid shape flattened on one side, much like a flattened, partially inflated football. It may
contain a partially or fully developed larva.
2.
Adult worms occasionally are seen in transparency tape preparations. The measure 1 cm long, have a
pointed tail posteriorly and transparent wings flanking the anterior end.
Reporting results:
A. Report her organism and stage. Do not use abbreviations.
Example: Enterobius vermicularis eggs present.
B. Report adult worms.
Example: Enterobius vermicularis adult worm present.
C. Report negatives.
Example: No Enterobius vermicularis eggs or adults seen.
Precautions
1.
The eggs of E. vermicularis are infectious if swallowed, so the person obtaining specimen must wear the
gloves.

2.

If opaque tape is submitted by mistake, a drop of immersion oil on the top of the tape will clear it enough to
proceed with the microscope examination.
Limitations of the test-cellulose tape preparation for pinworm examination

1.
2.

The female pinworm deposits eggs on the perianal skin only sporadically.
Without multiple tapes (taken on consecutive morning), it is not possible to determine if the patient is
positive, or negative for the infection

Anal swab method

A new anal swab concentration technique for detection of the eggs of Enterobius vermicularis consists of an
ordinary cotton swab, as used for throat cultures, dipped into a melted mixture of four parts of vaseline and one part
of Parowax and allowed to cool. The swab is put into a culture tube and given to the patient who brushes it lightly
over the skin surrounding the anus and then inserts it for about one quarter of an inch into the rectum. One swab is
used on each of two successive mornings upon awakening and replaced in the culture tube. The tubes are half filled
with xylol and allowed to stand until the wax coating is dissolved. The swab is removed and the tube centrifuged.
The supernatant fluid is carefully poured off and the sediment is then examined for eggs. This technique eliminates
discomfort and gives better results than the NIH swab. R.T.L

Hookworms

Examination of a stool sample

Complete blood count (CBC) with differential
-Blood tests to check for anemia and nutritional deficiencies
Hookworm infection diagnosis is made by identifying hookworm eggs in a sample of stool. Stool should be
examined within several hours after defecation.
Blood tests for anemia and nutritional deficiencies, particularly iron, are also done.
Diagnosis of established hookworm infections is made primarily by means of microscopic
identification of characteristic eggs in the stool (Fig. 4.5). In an infected human, a single
adult female hookworm will produce thousands of eggs per day. Because hookworm
infections will oft en not present with specifi c signs and symptoms, the clinician typically
requires some index of suspicion, such as local epidemiology or country of origin or travel, to
request a fecal examination for ova and parasites. Hookworm eggs are colorless and have a
single thin hyaline shell with blunted ends, ranging in size from 55-75 m by 36-40 m.
Several sensitive egg concentration techniques, such as the formalin-ethyl acetate
sedimentation method, can be used to detect even light infections. Where concentration
procedures are not available, a direct wet mount examination of the specimen is adequate
for detecting moderate to heavy infections. A single stool sample is often suff cient to
diagnose hookworm infection, with diagnostic yield not appreciably increased by examining
further specimens. Although examination of the eggs cannot distinguish between N.
americanus and A. duodenale, this is not clinically relevant. Diff erentiation between the two
species can be made by either rearing fi lariform larvae from a fecal sample smeared on a
moist fi lter paper strip for fi ve to seven days (the Harada-Mori technique) or by recovering
the adult worms following treatment and examining for such features as the mouthparts.
Besides microscopic examination of feces, eosinophilia is a common fi nding in persistent
infection and also during the phase of migration of larvae through the lungs. A chest
radiograph will usually be negative during the pulmonary phase of larval migration, although
sputum examination may reveal erythrocytes, eosinophils and rarely migrating larvae.
Dermatological infection with the zoonotic hookworms A. caninum, A. braziliensis and U.

stenocephala is primarily diagnosed clinically, although eosinophilia and elevated serum IgE
occur in between 20% to 40% of patients with cutaneous larva migrans. Since larvae do not
migrate to the gastrointestinal tract to develop into adult worms, fecal examination will be
negative in these cases.