Iman M.

Fawzy
Clinical Pathology MD. PhD
Mansoura, Egypt
Iman M. Fawzy
Clinical Pathology MD. PhD
Mansoura, Egypt
The best use of microbiologic lab. involves
sending only relevant specimens.

For obtaining good quality sample for
microbiologic examination, certain
precautions should be considered.
Rapid transporting of the
specimens to the lab.
Collection of
specimens before
antibiotic therapy.
Sample collection as
early as possible in
the disease.
Containers used for
collection should be
sterile, tightly capped,
and clearly labeled.
Cleaning the site
of infection
General precautions
Any specimen should be accompanied by request
containing:
Patient's name, age, gender, in or outpatient,
ward number, home area .

Type & source of specimen, date & time of
collection.

Investigation(s) required.

Clinical note summarizing the patient's illness,
suspected diagnosis, antimicrobial therapy.

Name of clinical officer .
Request form
Proper Sampling in Clinical Microbiology Laboratory
Collection of good quality specimens Depends on:

1. The optimal time of specimen collection .
2. The correct type of specimen
3. Well collected specimens with minimum
contamination from normal flora of the patient or
the person collecting the specimen.
4. Adequate amounts of each specimens and
appropriate no. of specimens
5. Clearly labeled safe specimens
Proper Sampling
Proper Sampling
Optimal time of collection of specimens: - 1
Specimens for the culture of bacteria must
be collected before the start of antibiotics
Blood films for malarial parasites are
usually best collected just as the patient's
temperature starts to rise
When infective endocarditis is suspected,
three blood culture sets can be collected
within a 24 h period irrespective of the
temperature of the patient .
Proper Sampling
Optimal time of collection of specimens: - 1
Serology is usually most satisfactory when
a fourfold or greater rising antibody titre
to a pathogen can be demonstrated in
paired sera.
Specimens for electron microscopy or
isolation of viruses are most likely to give
positive results when collected during the
most acute stages of the disease
Proper Sampling
Correct types of specimens - 2
Patients with possible bacterial meningitis
should have blood cultures as well as
cerebrospinal fluid collected.
Cervical, urethral and preferably rectal swabs should
be collected rather than high vaginal swabs from
female patients with suspected gonorrhoea.
Pernasal swabs should always be collected from
children who may have pertussis rather than a
throat or ordinary nose swab.
Proper Sampling
3- Well- collected specimens with minimal
contamination from normal flora
All staff should become aware of the importance of
minimizing the contamination of specimens.

Poor-quality specimens include saliva instead of
sputum or a salivary-mucoid sputum sample instead of
a mucopurulent specimen from a patient with
pneumonia .

Mid-stream specimens of urine to avoid excessive
contamination.
Proper Sampling
3- Well- collected specimens with minimal
contamination from normal flora
Throat swab collected should not touch the buccal
mucosa or tongue .

A high vaginal swab should be collected using a vaginal
speculum which should not be wet with antiseptic
solution with care not to touch the lower region or
perineum.

Blood or cerebrospinal fluid (CSF) cultures can be
contaminated by the skin flora of the patient or from
the doctor .
Proper Sampling
Adequate quantity of the specimens - 4
The collection of adequate quantities of early morning
sputum.

At least two samples of faeces on different days are
desirable from patients with diarrhoea.

When carriage of Salmonella typhi is investigated in a
food handler, 6 samples of faces should be cultured.



Proper Sampling
Adequate quantity of the specimens - 4
Patients with diarrhea at least 2 specimens of
faces is collected for culture of Salmonellae or
Shigella.

Serological investigations usually require paired sera.
Proper Sampling
Clearly labeled and safe containers - 5
Microbiological investigations should be placed in leak-
proof containers.

Each container should be enclosed in a plastic bag.

Examples of microbiological hazards to staff handling
leaking containers include TB, and viruses such as HBV
and HIV.
Urine Samples
Morning sample.

Washing the site with soap & water 3 times,
and in female from front to backward.

Mid stream sample for routine culture
& 24h (3 times) for T.B.

In children:
Suprapubic aspiration
Adhesive bag
Urine Samples
If the patients is already catheterized, catheter
sample can be used.

Sterile, dry, wide-necked, leak proof container is
used.

Collection of 10 20 ml urine.

To preserve the sample, (0.Ig /10ml urine) boric acid
is used.
Urine Samples
When gonorrhea is
suspected immediate
culture (bed side) or
using transport
medium
Blood Samples
During peak of fever.

Sterilizing the site of venipuncture
with alcohol 70% & iodine .

Samples for aerobic blood culture
are collected in the green capped
bottles, while those for anaerobic
blood culture are collected in the red
capped bottles.
Blood Samples
For adults at least, 5mL of blood are
collected

while for infants and children 2mL of
blood only are collected.

For automated blood culture system
special bottles are used.

Blood is introduced through the rubber
cap.
Blood Samples
Serum can be used for culture on fluid medium.

Blood for serological diagnosis is collected in screw
capped tubes without anticoagulant.
Body Fluids Samples
Sterilizing the site with alcohol 70% and iodine.

Rapid transporting to the lab especially for CSF.

Using wide mouth, sterile containers to which 0.3ml
of 20% Na citrate is added to prevent clot
formation.
Sputum Samples
Morning sample.

Washing the mouth with water and brushing the
teeth.

Sputum rather than saliva .

In infants, gastric lavage can be used.

Sputum Samples
An alternative method, the tongue is depressed
and cough is induced by touching the larynx,
material from the trachea expelled during the
cough will adhere on the swab.

Samples for T.B. are 3 consecutive morning
samples.
Swabs
It is a sample device

consist of sterile pledget of absorbent material,
usually cotton-wool or synthetic fiber, mounted on a
thin wire or stick.

Swabs for Bacterial (red) and
Viral (green) Cultures
Swabs
The absorbent material may be inhibitory to the
pathogens, some swabs may be treated to prevent
inhibitory effect:

Boiling the swab in phosphate buffer
Coating the swab with serum
Or coating the swab with albumin or charcoal
Swabs
Uses of swabs:
for taking specimens of exudates from nostrils ear,
skin, throat, wounds and other accessible lesions.

baby swabs: very fine swab.
Swabs
Uses of swabs:
pernasal swab: used for diagnosis of
whooping cough. It is passed along the
floor of nasal cavity to reach the
nasopharyx . It is small with flexible
wire .


post nasal swab: the terminal 20mm of
long stiff metal wire is bent at an
angle of 45 o to be introduced through
the mouth to reach nasopharynx.
Used for diagnosis of meningococcal
carrier.
Swabs
Uses of swabs:
Laryngeal swab : used for obtaining
sample from bronchial secretion for
diagnosis of TB. About 50 mm of the
wire is bent at an angle of 60 o .

High vaginal swab used fort diagnosis
of genorrhea and puerperal fever . It
is about 22cm long to reach
endocervix.
Throat Swabs
Washing the mouth with water.

Depression of the tongue with tongue depressor.

Sample from posterior pharynx or tonsils.

Swab should not touch the buccal mucosa.
Wounds
Clean the wound with sterile gauze soaked in
sterile saline.

Squeeze the wound.

Swab pus or squeezed exudates.
Eye Swabs
Washing of eyes by water and soap.

Apply swab from lower conjunctival fornix .
Ear Swabs
Clean the site with sterile saline.

Dry the area.

Apply swab to get specimen.
Stool Samples
Stool is collected in wide-mouth container.

Stool is collected in leak proof sterile container
into which the excreta can be passed directly.

Alternatively, rectal swab may be taken.
Stool Samples
For preserving Salmonella, place several drops of
luiquid faces or faecal emulsion on a sheet of
filter paper and allow it to dry and preserve it in a
cellophane envelop for transporting to the lab. In
the lab a saline suspension of material from the
filter paper is plated on a selective media.

For Cholera. The faeces is sent in bile peptone
medium to prevent the death of Vibrio cholera .
Gynacologicol sampling
Cervical swab is preferred rather than vaginal ones.

Dry, sterile speculum should be used or moistened
with sterile warm water.

No bimanual examination performed.
Gynacologicol sampling
Cleanse the cx. with swab moistened with sterile
saline.

Sterile swab is used for sample collection by
rotating it in the cervix.

When gonorrhea is suspected immediate culture
(bed side) or using transport medium .
Sampling for Mycologic exam
Clean the site with alcohol 70%.

Sterile scalpel is used for scrapping.

Sterile scissor is used for trimming the
nails.

Scales are transferred in clean dark
paper.

For systemic mycosis, precautions are
those for blood culture.
Sampling for Clinical Virology
Collection of specimen during most active
stage of the disease.

Samples as urine , stool , CSF , sputum are
transported directly to the lab while swabs
from throat , nose vesicles , cervix are
transported in VTM.

If delay is in due, keep specimens at 4
0
C.

Biohazard label should be used for
containers of HBV, HCV and HIV.