Microbiology Sample Collection

Sample Collection &
transport
Objectives
• General consideration for sample
collection
• Sample safety considerations
• Rejection criteria
• Collection of different samples from
different sites
• Transportation
• Reference
General consideration for proper
sample collection
-Every laboratory should provide proper

guidelines for collection of samples
-All diagnostic information depends on

quality of sample recieved
-If sample collection, transport, media are

not proper, it will give false results
• Collect sample before administering
antimicrobial agents when possible.
• Collect sample with as little

contamination from indigenous
microbiota as possible to ensure that
the sample will be representative of the
infected site.
• Utilize appropriate collection devices.
Use sterile equipment and aseptic
technique to collect specimens to
prevent introduction of microorganisms
during invasive procedures.
• Clearly label the specimen container

with the patient’s name and
identification number. Always include
date and time of collection and your
initials.
• Collect an adequate amount of
specimen. Inadequate amounts of
specimen may yield false-negative
results.
• Identify the specimen source

and/or specific site correctly so
that proper culture media will be
selected during processing the
laboratory.
• Collect specimens in sturdy, sterile,
screw-cap, leak proof containers with
lids that do not create an aerosol when
opened.
• Collect sample after proper preparation

of area by spirit and povidone iodine
Sample Safety considerations
• Follow universal precaution guidelines. Treat

all specimens as potentially biohazardous.
• Laboratory workers should use appropriate

barrier protection (such as gloves and
laboratory coat or gown) when collecting or
handling specimens. If splashing may occur,
protective eyewear, face masks, and aprons
may be necessary.
• Do not contaminate the external
surface of the collection container
and/or its accompanying paperwork.
• Minimize direct handling of specimens

in transit from the patient to the
laboratory. Use plastic sealable bags
with a separate pouch for the
laboratory requisition orders or
transport carriers (for example, small
buckets with rigid handles).
Label High risk Specimens
• Sputum with
suspected
Tuberculosis
• Fecal samples
suspected with
Cholera, Typhoid,
• Serum when
suspected with
HIV/ HBV/HCV,
infections
REJECTION CRITERIA
• Leaking/broken container
• Insufficient amount
• Improper labelling
• More time lag between collection and
transport
• Improper transport media
• Improper transport temperature
• Hemolysed sample
Important questions before
collecting a specimen
• Are you suspecting an Infection ?
• If so what is the Nature of infection,
eg Bacterial, Viral, Mycological or
Parasitological
• Which tests are your priority ?
• When to collect the specimen ?
• How to collect the specimen ?
• Am I choosing the correct container ?
• Why to send the specimens promptly,
if not what I should do ?
A Request form
• Should include:
-Name ,Age ,Sex,Address

-IPD/ OPD No ,ward,
-Time and Date,
-Urgent / Routine,
-Type of specimen
-Investigation needed
-History of patient
-Provisional diagnosis
-Doctor’s name and sign
When to Request Transport
Medium
• When facilities are not available to perform
the desired tests at the place of collection or
laboratory located far away, request the
Diagnostic laboratories to advice on
transportation of specimens, and consider how
to preserve and transport in ideal medium
before it is processed
• Popularly used transport medium are

-Amie 's transport Medium:
Gonococcal infection
-Carry blair medium: stool
-V.R Medium: Stool
Collection of different samples
• Blood
• Urine
• Stool
• Rectal swab
• Sputum
• Csf
• Mycology samples
• Wound swabs
• Water sample
Blood
• Take proper precaution (gloves)
• Avoid contamination
• Palpate vein
• Apply disinfectant
• Use sterile needle and syringe
• Collect 5-10 ml blood for two sets of culture
each
• In children collect 2-5 ml
• Infuse it into BHI after cleaning with spirit
Blood for serology
• Same precautions
and method
• Collect blood into
plain vacutte
• It will coagulate
• Centrifuge it at
1500-2000 rpm for
5 mins
• Thus serum will be
separated
Urine
For females
• Rinse area with soap and water
• Clean it with sterile gauze piece
• With both labia apart allow first few ml
of urine to drain
• Take mid stream clean catch urine
• This will prevent contamination
• Collect it into wide,sterile,leak proof
container
• Transport it directly to the laboratory
after proper labelling
• If delay in transport, preserve it at 4-6
degree centigrade
• 10000-100000 cfu/ml will show
contamination mostly
For males
• Wash hands and retract
prepuce before collecting
sample
• If it is not possible to
obtain urine, suprapubic
aspiration or
catheterization may be
used
• Catheterised urine
should not be used
ideally bcz of presence
of bacteria in urethra
• For that first apply
disinfectant over
infusion port and then
collect urine
Stool
• Collect it into wide mouth,clean,
sterile,leak proof container
• Label properly
• Min 5 gms is needed if solid stool
• And 2 ml if liquid stool
• It should not be contaminated with
urine
• If not possible to collect it as in
children, elderly, debilitated patients–
collect rectal swab
• Do not referigerate stool
• If delay in transport preserve it into
10% formalin, buffered glycerol saline
Rectal swab
• Take sterile swab
• Apply it in anal canal
• Rotate it for 10 seconds
• Avoid contact with skin
• Seal it directly into swab container to
prevent contamination
• Transport immediately to lab , if not
possible,preserve at 4-6 degree
centigrade
Sputum
• Wide container of 50-100 ml capacity
• For M.TB two samples are collected
• 1st at on the spot and 2nd at next day
morning
• Morning sample is more reliable bcz of
colonization of bacteria in LRT at night
• Before this ask patient to gargle with
normal saline and take deep breath and
then take sample in one bout
• This will prevent contamination
• There should not be saliva or liquid
• If there are >25 epithelial cell per LPF,
sample is rejected as it shows
contamination
• It should be collected before antibiotic
treatment has started
Procedure to collect CSF
• Collect only 3-5 ml into

a labeled sterile
container
• Removal of large volume
of CSF lead to
headache,
• The fluid to be
collected at the rate of
4-5 drops per second.
• If sudden removal of
fluid is allowed may
draw down
cerebellum into the
Foramen magnum and
compress the
Medulla of the Brain
Preservation of CSF
• It is important when
there is delay in
transportation of
specimens to
Laboratory do not
keep in Refrigerator,
which tends to kill H.
Influenza
• If delay is
anticipated leave at
Room Temperature.
MYCOLOGY SAMPLES
• Clean site with 70% ethanol to help eliminate
surface contaminants. Using a scalpel, skin
scrapings should be made from the active
periphery of the lesion. For nails Scrapings
should be deep enough to assure acquiring
recently invaded tissue Submit scrapings in a
sterile Petri dish or container.
• Hair - Use forceps to pluck involved hairs
from the edges of the patches. Submit hair,
including shaft, in a sterile Petri dish or
container.
• Other - Collect and submit specimens as
described for specific type. Specimens
associated with the systemic and deep
seated mycoses are obtained from a wide
variety of sources. They should be obtained,
whenever possible, under aseptic conditions
and in sufficient quantity for both
microscopic and cultural examinations.
Samples from wounds
• The ideal sample is pus or exudates
should be submitted in a small screw-
capped bottle in firmly stoppered tube
or syringe or a sealed capillary tube.
• Fragments of excised tissue removed at

wound toilet or curettings from
infected sinuses and other tissues
should be sent in a sterile container.
• The swabs are inefficient sampling
device and tends to desiccate the
specimen and trap the bacteria which
are then not released on to culture
plate
Water for bacteriology
• Water course or reservoir - collect from a

depth of at least 20 cm
• Dug well - do not allow the bottle to touch
the sides of the well
•Collection
At least 200 ml of water sample from
the source
• In sterile glass bottles OR autoclavable
plastic bottles
• tight screw capped lid
• securely fitting stoppers/caps
• an overhanging rim
• Handling and transportation
Test the water sample within 3
hours of collection
• keep at ambient temperature
• If delayed:
• pack sample on ice
• test refrigerated sample within 24 hours
Transportation of samples
• Stool, csf and sputum should be
transported at room temperature, not in
refrigerator
• Urine , swabs, skin samples,water & food
samples are transported asap at room
temp,but if it is not possible, preserve
them in refrigerator
References
• Koneman’s color atlas and Textbook of
diagnostic microbiology,sixth
addition,chapter 2-guidelines for
collection,page-74,76,81,83,93,100
• Mackie & McCartney:Practical Medical
Microbiology,14th edition,chapter:5,Sample
collection:page-95-96
• IDSP guidelines-
www.idsp.nic.in/idsp/IDSP_2WeekCourse.../
WorkLaboIDSP.ppt
• NACO guidelines-
naco.gov.in/upload/Blood%20Saftey/Sample%20
Trtansport.pdf
• Role of specimen collection in infectious disease
by Dr. T. V. Rao MD
• WHO guidelines 2011
Thank You

Microbiology Sample Collection

Uploaded by

Copyright:

Available Formats

Microbiology Sample Collection

Uploaded by

Document Information

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

Microbiology Sample Collection

Uploaded by

Copyright:

Available Formats

Sample Collection &

-Every laboratory should provide proper

-All diagnostic information depends on

-If sample collection, transport, media are

• Collect sample with as little

• Clearly label the specimen container

• Identify the specimen source

• Collect sample after proper preparation

• Follow universal precaution guidelines. Treat

• Laboratory workers should use appropriate

• Minimize direct handling of specimens

-Name ,Age ,Sex,Address

• Popularly used transport medium are

• Collect only 3-5 ml into

• Fragments of excised tissue removed at

• Water course or reservoir - collect from a

You might also like