MALAMULO COLLEGE OF HEATH SCIENCES

DEPARTMENT OF BIOMEDICAL SCIENCES

SUBMITTED TO : LINLY LINJE

SUBMITTED BY : MATTHEWS NJOBVU(BMS/22/694)
PROGRAMME : DIPLOMMA IN BIOMEDICAL
SCIENCE
COARSENAME : MEDICAL PARASITOLOGY
YEAR : TWO
TOPIC : SPECIMENS COLLEC
 SPECIMENS COLLECTION

1. BLOOD

Collection time

 Before the start of antibiotic therapy. If this is not possible due to seriousness of the
patient’s illness, two sets of blood should be drawn from two different sites before the
antibiotic is administered

Equipment

 Request form, labels, and blood culture bottles for aerobic and anaerobic cultivation,
antiseptic solution, cotton wool sponges, needles, syringes and tourniquet.

Procedure

 Fill in request form, identify the patient and explain what will happen.
 Collect and assemble the equipment. Remove the protective cap from the culture bottles,
if present.
 Apply tourniquet and select an appropriate vein for the collection, usually the antecubital
vein.
 Disinfect the skin: apply the an antiseptic solution to a cotton wool sponge and rub a 5cm
square area around the selected site for one minute. Leave the antiseptic to dry and
discard the sponge. Make a new sponge and cleanse the site again, beginning at the
Centre and scrubbing in a circular motion outwards. Let it dry.
 If necessary, palpate the site before the venipuncture but do not touch the prepared site
with the fingers that are not disinfected.
 Insert the needle of the syringe into the vein and draw the volume needed for the culture
or test.
 Remove the tourniquet and withdraw the needle from the vein
 Immediately apply pressure to the punctured with a clean cotton sponge.
 Inoculate the culture bottles carefully, adding the correct amount of blood. Be careful that
no air is injected.
 Put a label on the bottle, indicating the patient’s identification, ward number, and time of
collection, so as not to cover the area occupied by the medium.

Instructions or safety precautions

 Do not puncture the same sire twice as this may cause infection.
 Blood from the patient is potentially infected with hepatitis or AIDS, so when injecting it
in the culture bottles so be careful not to prick your fingers.
 Needles should not be recapped, but discarded in a safety container.
Storage

 According to the manufacturer’s instructions. Do not store inoculated bottles in the

refrigerator.

Transportation

 Bring the blood culture bottles to the laboratory immediately, and place the culture
bottles immediately in the incubator.

Testing

 Blood smear; this test is used to look for parasites that are found in the blood. By looking
at a blood smear under a microscope, parasitic diseases such as filariasis, malaria, or
babesiosis, can be diagnosed.

 This test is done by placing a drop of blood on a microscope slide.

Reporting

 In case of positive culture, the result should be conveyed immediately to the treating
physician or doctor no duty.
 Negative results are only reported after 7 days of incubation.
 If slow growing organisms are suspected-Brucella spp,, Francisella tularensis- it should
be clearly indicated on the requisition form, and the culture bottles should be further
incubated for another 1 to 2 weeks before being reported out as negative.

2. STOOL FOR BACTERIA

Test material

 Diarrheal stool.
 Formed stool should be rejected for bacterial examination but for parasites is
recommended.

Collection time

 Fecal specimen should be collected in the early stages of the disease when pathogens are
usually present in the stool in the large numbers, and preferably before antibiotic
treatment is begun.
 The specimen should be collected in the morning to reach the laboratory before noon, so
that it can be processed the same day.
Equipment

 A suitable specimen container provided with a lid: a clean glass cup, a plastic or a waxed
cardboard box, or a special container with a spoon sponge to the lid or stopper, two sticks
to transfer the specimen to the container. The use of the penicillin bottles, match boxes
and banana leaves, should be discouraged as they expose the laboratory staff to risk of
infection.

Procedure

 Instruct the patient on how the specimen should be collected and transferred to the
container; provide him/her with sticks and containers.
 The stool should be collected on a piece of toilet tissue or old newspaper.
 A sample is transferred with the sticks to the container. The specimen should contain at
least 5g of feaces and, if present, those parts that contain blood and/or mucopus should be
selected. The specimen should not be contaminated with urine. Close the lid.
 The specimen should be transferred to the laboratory and processed within a few hours.
 If delay, in shipment of the specimen to the laboratory is anticipated, stool specimens
should be placed in a container with transport medium (Cary-Blair, Stuart or Amies) or
glycerol-phosphate buffer 0.033M.

Storage

 Refrigerated (2-8 o C).

 For parasites, refrigeration is not recommended.

Transportation

 Preferably in a cooling box (2-8 oC). If shipped through the mail, the transport tubes
should be carefully wrapped and packed in a suitable, durable container that does not
break during shipment.

Testing

 Wet preparations of fresh stool (<2 h since defecation) can be scrutinized microscopically
to visualize the motile trophozoites of Giardia and other ameboid parasites.
 Stained preparations should always be examined even if direct preparations are positive,
as additional parasites may be detected

Reporting

 Negative results are sent out 48 hours after receipt of the specimen.
 Results of positive cultures can be expected in 2-3 days.
 3. WOUND SWAB

Test material

 Purulent material or tissue

Equipment

 Swabs: Forceps, cotton wool sponges, sterile cup with sterile saline, sterile cotton wool
swab, test-tube with transport medium, and bandage.

Aspiration

 Forceps, cotton wool sponges, 70% alcohol, syringe, needle, sterile cotton wool swab,
test-tube with transport medium, bandage.

Procedure

 Inform the patient.

 No-touch technique: remove bandage with the forceps.
 With the forceps take a sponge, dip it in the saline and wash the surface of the wound free
from exudate.
 Remove the swab from its covering and extend the tip of the swab deep into the wound,
taking care not to touch the adjacent skin margins.
 Remove the stopper from the test-tube with transport medium, plunge the swab into the
transport medium and replace the stopper. It the wooden stick of the swab is too long,
break off the end over the rim of the test-tube.
 Apply new bandage.
 Wash hands and fill in the request form.

Aspiration

 Inform the patient

 With the forceps take a sponge, wet it with alcohol and decontaminate the margins of the
wound. Let the alcohol dry in between, and repeat the decontamination once more.
 Insert the needle through the decontaminated margin and aspirate the material from the
depth of the wound.
 Squirt some of the material slowly on to a sterile cotton wool swab and plunge the swab
into the transport medium. The rest of the material is deposited on a microscope slide and
a smear is made with the needle.
 Apply new bandage
 Wash hands and fill in the request form.
Storage

 Refrigerated(2-8 oC)

Transportation

 In cooling box (2-8oC), preferably.

Testing

 Semi quantitative swab culture:  Some bacteria infecting a wound may require air for
growth (aerobic) while some require a no-oxygen or reduced-oxygen environment
(anaerobic or microaerophilic)
 However, there is research which shows a correlation between bacteria identified in a
swab culture and the bacteria identified from a tissue sample, specifically a biopsy.
 A swab culture will not give the level of quantitative results as a biopsy. However, if
done appropriately, the results will identify the microbes that need to be targeted
with antimicrobial interventions.

Reporting

 A preliminary result is reported 2 days after the specimen is received. The final report is
often delayed for 4-5 days.

4. TISSUE BIOPSY

Specimen

 Tissue biopsy removed by surgery

Collection time

 Preferably before the patient is treated with antibiotics

Equipment

 Mask, gown, gloves, sterile forceps, sterile specimen container.

Procedure

 Divide large tissues into small ones(0.5cm) in diameter

 With aseptic technique transfer the specimen to a sterile container and immediately bring
it to the laboratory.
  If transportation is prolonged for more than an hour, place the biopsy in a tube with
transport medium and push it to bottom of the agar with a sterile swab.
 Fill in a request form and label the container with patient’s identification.

Storage

 Refrigeration (2-8oC).

Transportation

 Preferably in cooling box (2-8oC) if transportation time is delayed for more than an hour.

Testing

 Routinely stained hematoxylin and eosin sections are suitable for examination for
parasitic agents, and many times, an accurate identification of the parasite can be
established in tissue sections. Occasionally, additional sections are required to further the
diagnosis.

Reporting

 Negative results are reported two days after the specimen is received, positive findings
may take from two to five days or even longer.

5. URINE

Test material

 Clean-catch, midstream urine specimen

Collection time

 For urine culture morning urine is recommended or, in case this is not possible, then a
specimen collected 2 hours after last mictutation.
 For the demonstration of eggs of Schistosoma heamatobium, a random urine sample
should be collected preferably in a petri dish between 10 a.m. and 2 a.m., as the
concentration of eggs is greater during this period, particularly in the last drops of the
passed urine.
 Exercise just before collection will increase the excretion of eggs.

Equipment

 A clean, preferably sterile container (50ml or more) , cotton wool or gauze sponges, soap,
handwarm water, and bedpan.
Procedure (for bacterial and viral investigation)

 Give the patient a suitable container.

 Instruct the patient before the collection, preferably with illustration.
 Tell the patient not to touch inside or rim of the container.

Male

 If not circumcised, draw back the foreskin to avoid contaminating the urine.
 Begin to urinate, but pass the first portion into the toilet.
 Collect the mid-portion of urine into the container, and pass the excess into the toilet.

Female

 Squat over the toilet and separate the labia with one hand
 Void the first portion of urine into the toilet
 Collect the mid-portion of urine into the container and pass the excess into the toilet.

Processing urine for eggs of Schistosoma heamatobium

 Shake the bottle containing urine and fill a conical specimen glass with urine
 Let the urine in the specimen glass stand for at least 30 minutes
 By that time any Schistome eggs will have settled at the bottom of the specimen glass.
Without disturbing the deposit, carefully pour away the supernatant urine so that only
10ml are left
 Mix up the deposit by shaking and pour this into a specimen tube.
 Do the laboratory test

Important

 It is important that the urine is collected ‘’clean’’, as any discharge or pus from the
vagina or external genital added to the urine will invalidate the examination.

Storage

 All urine specimens should be brought to the laboratory within an hour of collection. If
this is not possible, the urine sample should be refrigerated just after collection and then
be brought to the laboratory to be processed within 24 hours.

Transportation

 In a cooling box (2-8oC) except when the transport time is very short.

Testing
  The definitive diagnosis of urinary schistosomiasis (Schistosoma haematobium) is
established by demonstration of S. haematobium eggs in urine.

 The specimen should be immediately centrifuged at 400 × g and the sediment examined
by wet mount.

 Trichomonas vaginalis motile trophozoites may also be found in the urine, especially in

infected male patients.

 To look for the presence of trophozoites, the urine specimen should be centrifuged at 400
× g, the sediment mixed with a drop or two of saline, and examined by wet mount.
Temporary stains, such as methylene blue or malachite green, are also helpful

Reporting

 Negative bacterial cultures are reported 1 day after receipt of specimen and positive
cultures after 1-2 days viral culture may take from 5-7 days.
References

Garcia, L. S. (2006). Diagnostic Medical Parasitology.

Lawrence R . Ash, T. C. (2008). Parasites, a guide to Laboratory Procedures and identification . America :
American Society of Clinical Pathologists.

Pathol., J. C. ( 2003). Journal of Clinical Pathology . Best Practice No 174 - PMC.

Zeibig, E. (2012). Clinical Parasitology: A Practical Approach. Elsevier Health Sciences.