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COLLECTION OF BLOOD FOR
CULTURING
Dr.T.V.Rao MD

Dr.T.V.Rao MD

1
What is a Blood Culture?
• A blood culture is a
laboratory test in which
blood is injected into
bottles with culture
media to determine
whether
microorganisms have
invaded the patient’s
bloodstream.
Dr.T.V.Rao MD

2
Need for Blood Culture?
No microbiological test is more essential to the

clinician than the blood culture. The finding of
pathogenic microorganisms in a patient’s
bloodstream is of great importance in terms of
diagnosis, prognosis, and therapy.”
- L. Barth Reller, Clin. Infect. Diseases, 1996

Dr.T.V.Rao MD

3
Blood Culture is done to Detect
Infectious Diseases
• Blood culture is a
microbiological culture of
blood. It is employed to
detect infections that are
spreading through the
bloodstream (such as
bacteremia, septicemia
amongst others). This is
possible because the
bloodstream is usually a
sterile environment
Dr.T.V.Rao MD

4
Blood culturing most important
and life saving Investigation
Needs optimal Methods for
Diagnosis of Blood Borne Pathogens

Dr.T.V.Rao MD

5
Blood Collection
• Aseptic collection
procedure is critical
Amount of blood
– 1:10 ratio of blood to
broth
– Younger than 10 years –
1 ml of blood for every
year of life
– Over 10 years – 20 ml

Dr.T.V.Rao MD

6
Blood Collection
• Frequency of Collection
– Depends if bacteremia is
transient, intermediate
or continuous
– Number of cultures
collected are usually
inversely related to the
type of bacteremia
– Usually x3 from different
body sites

Dr.T.V.Rao MD

7
Blood Culture Methods
• Conventional Broth Systems

–One aerobic bottle and one
anaerobic bottle per blood
collection
–Aerobic broth contains soybean
casein digest broth, Tryptic or
trypticase soy broth, Brucella agar
or Columbia broth base
–Anaerobic broth is usually the same
as aerobic with addition of 0.5%
cysteine in an aerobic environment
Dr.T.V.Rao MD

8
Venipuncture
• Venipuncture is the process of obtaining
intravenous access for the purpose of
intravenous therapy or obtaining a sample of
venous blood. This procedure is performed by
medical laboratory scientists, medical
practitioners, some EMTs, paramedics
phlebotomists and other nursing staff.
Venipuncture is one of the most routinely
performed invasive procedures and is carried
out for two reasons, to obtain blood for
diagnostic purposes or to monitor levels of
blood components (Lavery & Ingram 2005).
Dr.T.V.Rao MD

9
Phlebotomy Definition
• phle·boto·my (fli) noun
the act or practice of
bloodletting as a
therapeutic measure
• Phlebotomy from Greek
words, phlebo, relates
to veins, tomy, relates
to cutting.
• Opening a vein to
collect blood
Dr.T.V.Rao MD

10
LABELING THE SAMPLE
– Properly labelled sample is essential so that
the results of the test match the patient. The
key elements in labelling are:
• Patient's surname, first and middle.
• Patient's ID number.
• NOTE: Both of the above MUST match the
same on the requisition form.
• Date, time and initials of the phlebotomist
must be on the label of EACH tube.
Dr.T.V.Rao MD

11
Principles for Collection
• Gloves will be worn in accordance with standard
precautions.
• •Appropriate verification of the patient's
identity, by means of an armband or area specific
procedure, will occur before the specimen
collection.
• •Cultures should be drawn before
administration of antibiotics, if possible. ???
• Blood cultures should not be drawn from
lines, but should be drawn viavenipuncture.
Dr.T.V.Rao MD

12
What Materials We need
•
•
•
•
•

•
•
•
•
•
•

Chlorhexidine swabs (1-2 packages)
Alcohol swabs
Blood culture bottles (2 bottles per set)
2 syringes (adult: 20 cc, paediatric: 5 cc)
2 needles (adult: 22 gauge or preferably larger butterfly
or standard needle; pediatric: 25 or 23 gauge butterfly or
standard needle)
Gloves (sterile &nonsterile)
Tourniquet
Sterile gauze pad
Adhesive strip or tape
Self-sticking patient labels
Plastic zip lock specimen bags
Dr.T.V.Rao MD

13
The requisitions form should be completely filled
out, and the requisition must indicate the tests
ordered.

Dr.T.V.Rao MD

14
Self Protection
A few ways to make sure your
role in the collection process is
carried out with
efficiency, orderliness and safety

Dr.T.V.Rao MD

15
Steps 1 – 3, Check, Explain, Wash
• 1.Identify the patient
• 2.Explain the
procedure to the
patient.
• 3.Wash hands with
soap and water with
friction for 15 seconds
or use alcohol based
hand rub
Dr.T.V.Rao MD

16
Materials
•
•
•
•
•

•
•
•
•
•
•

Chlorhexidine swabs (1-2 packages)
Alcohol swabs
Blood culture bottles (2 bottles per set)
2 syringes (adult: 20 cc, paediatric: 5 cc)
2 needles (adult: 22 gauge or preferably larger butterfly or
standard needle; pediatric: 25 or 23 gauge butterfly or
standard needle)
Gloves (sterile &nonsterile)
Tourniquet
Sterile gauze pad
Adhesive strip or tape
Self-sticking patient labels
Plastic zip lock specimen bags
Dr.T.V.Rao MD

17
. Barrier protection for the phlebotomist

consists of the latex gloves.

Dr.T.V.Rao MD

18
Obtaining Blood
• Locate the vein
• Prep kit
– Alcohol 5 sec. Dry 30-60 sec ( resource poor conditions )
– Ideal to collect with alcohol swabs containing 2% Chlorhexidine
and 70% isopropyl alcohol

• Remove caps, clean with alcohol
• Put on gloves
• Without palpating, draw 20 ml and put 10
in anaerobic and 10 in aerobic bottle
• Dispose of syringe in sharps container
• Label bottles and send to lab
Dr.T.V.Rao MD

19
Method of Blood Collection
•

A minimum of 10 ml of blood is taken

through venipuncture and injected
into two or more "blood bottles"
with specific media for aerobic and
anaerobic organisms.
• The blood is collected using clean
technique. This requires that both
the tops of the culture bottles and
the venipuncture site of the
patient are cleaned prior to
collection with alcohol swabs
containing 2% Chlorhexidine and
70% isopropyl alcohol.

Dr.T.V.Rao MD

20
The area of skin is cleaned with a
disinfectant, or an alcohol swab.
• Using sterile gloves, do not
wipe away the surgical
solution, touch the puncture
site, or in any way compromise
the sterile process. It is vital
that the procedure is
performed in as sterile a
manner as possible as the
persistent presence of skin
commensals in blood cultures
could indicate endocarditis but
they are most often found as
contaminants

Dr.T.V.Rao MD

21
The vein is anchored and the needle
is inserted.

Dr.T.V.Rao MD

22
The vacutainer tube is depressed into the
needle to begin drawing blood

Dr.T.V.Rao MD

23
Additional vacutainer tubes can be utilized. Determine what tests are
ordered and what tubes will be necessary BEFORE you begin to draw
blood, and determine the order of draw for the tubes. .

Dr.T.V.Rao MD

24
When the final tube is being drawn, release the tourniquet.
Then remove the tube, and remove the needle.

Dr.T.V.Rao MD

25
After the needle is removed from the vein, apply firm pressure
over the site to achieve haemostasis.

Dr.T.V.Rao MD

26
Apply a bandage to the area.

Dr.T.V.Rao MD

27
Preparation of Cap before Injecting
Blood
• Prep the rubber
cap of the blood
culture bottles
with an alcohol
pad in a circular
motion. Allow
the alcohol to
dry.
Dr.T.V.Rao MD

28
Inject the Blood …..
• Inject the blood
into the Selected
Media
• Gently rotate the
bottles to mix
the blood & the
broth (do not
shake
vigorously).
Dr.T.V.Rao MD

29
Follow the universal precautions when
disposing Needle

• Dispose of
needle in sharps
container and
dispose of other
waste in proper
container

Dr.T.V.Rao MD

30
Label the tubes, checking the requisition for the proper
identification.

Dr.T.V.Rao MD

31
Give the all possible Medical
Information
•
•
•
•
•
•
•
•

Patient’s name
• Hospital number (Patient ID)
• Patient’s location (room and bed #)
• Date and time of collection
• Collector’s initials
• Site of venipuncture
• Or other information as per facility
Include you Mobile Contact No – A vital
information can be delivered any time
Dr.T.V.Rao MD

32
Document the Medical Records
• Document the
following in the
medical record:
• –Date & time
specimen obtained
• –Site of specimen
collection
Dr.T.V.Rao MD

33
Frequency of Collection
• Frequency of Collection
– Depends if bacteremia is
transient, intermediate or
continuous
– Number of cultures
collected are usually
inversely related to the
type of bacteremia
– Usually x3 from different
body sites

Dr.T.V.Rao MD

34
Second Set
• If 2 or more sets
of blood cultures
have been
ordered, obtain
the second set in
the same manner
as the
first, making a
new venipuncture
at a different site.
Dr.T.V.Rao MD

35
Newer Blood Culture Methods
• Newer Blood Culture Systems
– Biphasic Broth-Slide System
• Agar “paddles” attached to top of bottle
• Closed system

– Continuous Monitoring Blood Culture Systems
•
•
•
•

BacTec – measures 14CO2
BacTec 9000 Series – measures CO2
ESP – measures consumption of gases
BacT-Alert – measures change in pH

Dr.T.V.Rao MD

36
The Contaminated Blood Culture
• If the skin is not adequately cleansed before
drawing blood for culture, bacteria on the skin
will be injected into the bottle, producing a false
positive blood culture
• It is difficult for the physician to determine
whether the bacteria growing in the blood
culture is a real pathogen causing bloodstream
infection or whether bacteria on the skin have
contaminated the culture. This can lead to
excess use of antibiotics and prolongation of
hospital stay.
Dr.T.V.Rao MD

37
Visit me for more articles of interest in
Medicine and Health care please visit…

Dr.T.V.Rao MD

38
• The programme created by Dr.T.V.Rao
MD as Technical Series for
Microbiologists in the Developing World
• Email
• doctortvrao@gmail.com

Dr.T.V.Rao MD

39

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Collection of blood for culturing

  • 1. COLLECTION OF BLOOD FOR CULTURING Dr.T.V.Rao MD Dr.T.V.Rao MD 1
  • 2. What is a Blood Culture? • A blood culture is a laboratory test in which blood is injected into bottles with culture media to determine whether microorganisms have invaded the patient’s bloodstream. Dr.T.V.Rao MD 2
  • 3. Need for Blood Culture? No microbiological test is more essential to the clinician than the blood culture. The finding of pathogenic microorganisms in a patient’s bloodstream is of great importance in terms of diagnosis, prognosis, and therapy.” - L. Barth Reller, Clin. Infect. Diseases, 1996 Dr.T.V.Rao MD 3
  • 4. Blood Culture is done to Detect Infectious Diseases • Blood culture is a microbiological culture of blood. It is employed to detect infections that are spreading through the bloodstream (such as bacteremia, septicemia amongst others). This is possible because the bloodstream is usually a sterile environment Dr.T.V.Rao MD 4
  • 5. Blood culturing most important and life saving Investigation Needs optimal Methods for Diagnosis of Blood Borne Pathogens Dr.T.V.Rao MD 5
  • 6. Blood Collection • Aseptic collection procedure is critical Amount of blood – 1:10 ratio of blood to broth – Younger than 10 years – 1 ml of blood for every year of life – Over 10 years – 20 ml Dr.T.V.Rao MD 6
  • 7. Blood Collection • Frequency of Collection – Depends if bacteremia is transient, intermediate or continuous – Number of cultures collected are usually inversely related to the type of bacteremia – Usually x3 from different body sites Dr.T.V.Rao MD 7
  • 8. Blood Culture Methods • Conventional Broth Systems –One aerobic bottle and one anaerobic bottle per blood collection –Aerobic broth contains soybean casein digest broth, Tryptic or trypticase soy broth, Brucella agar or Columbia broth base –Anaerobic broth is usually the same as aerobic with addition of 0.5% cysteine in an aerobic environment Dr.T.V.Rao MD 8
  • 9. Venipuncture • Venipuncture is the process of obtaining intravenous access for the purpose of intravenous therapy or obtaining a sample of venous blood. This procedure is performed by medical laboratory scientists, medical practitioners, some EMTs, paramedics phlebotomists and other nursing staff. Venipuncture is one of the most routinely performed invasive procedures and is carried out for two reasons, to obtain blood for diagnostic purposes or to monitor levels of blood components (Lavery & Ingram 2005). Dr.T.V.Rao MD 9
  • 10. Phlebotomy Definition • phle·boto·my (fli) noun the act or practice of bloodletting as a therapeutic measure • Phlebotomy from Greek words, phlebo, relates to veins, tomy, relates to cutting. • Opening a vein to collect blood Dr.T.V.Rao MD 10
  • 11. LABELING THE SAMPLE – Properly labelled sample is essential so that the results of the test match the patient. The key elements in labelling are: • Patient's surname, first and middle. • Patient's ID number. • NOTE: Both of the above MUST match the same on the requisition form. • Date, time and initials of the phlebotomist must be on the label of EACH tube. Dr.T.V.Rao MD 11
  • 12. Principles for Collection • Gloves will be worn in accordance with standard precautions. • •Appropriate verification of the patient's identity, by means of an armband or area specific procedure, will occur before the specimen collection. • •Cultures should be drawn before administration of antibiotics, if possible. ??? • Blood cultures should not be drawn from lines, but should be drawn viavenipuncture. Dr.T.V.Rao MD 12
  • 13. What Materials We need • • • • • • • • • • • Chlorhexidine swabs (1-2 packages) Alcohol swabs Blood culture bottles (2 bottles per set) 2 syringes (adult: 20 cc, paediatric: 5 cc) 2 needles (adult: 22 gauge or preferably larger butterfly or standard needle; pediatric: 25 or 23 gauge butterfly or standard needle) Gloves (sterile &nonsterile) Tourniquet Sterile gauze pad Adhesive strip or tape Self-sticking patient labels Plastic zip lock specimen bags Dr.T.V.Rao MD 13
  • 14. The requisitions form should be completely filled out, and the requisition must indicate the tests ordered. Dr.T.V.Rao MD 14
  • 15. Self Protection A few ways to make sure your role in the collection process is carried out with efficiency, orderliness and safety Dr.T.V.Rao MD 15
  • 16. Steps 1 – 3, Check, Explain, Wash • 1.Identify the patient • 2.Explain the procedure to the patient. • 3.Wash hands with soap and water with friction for 15 seconds or use alcohol based hand rub Dr.T.V.Rao MD 16
  • 17. Materials • • • • • • • • • • • Chlorhexidine swabs (1-2 packages) Alcohol swabs Blood culture bottles (2 bottles per set) 2 syringes (adult: 20 cc, paediatric: 5 cc) 2 needles (adult: 22 gauge or preferably larger butterfly or standard needle; pediatric: 25 or 23 gauge butterfly or standard needle) Gloves (sterile &nonsterile) Tourniquet Sterile gauze pad Adhesive strip or tape Self-sticking patient labels Plastic zip lock specimen bags Dr.T.V.Rao MD 17
  • 18. . Barrier protection for the phlebotomist consists of the latex gloves. Dr.T.V.Rao MD 18
  • 19. Obtaining Blood • Locate the vein • Prep kit – Alcohol 5 sec. Dry 30-60 sec ( resource poor conditions ) – Ideal to collect with alcohol swabs containing 2% Chlorhexidine and 70% isopropyl alcohol • Remove caps, clean with alcohol • Put on gloves • Without palpating, draw 20 ml and put 10 in anaerobic and 10 in aerobic bottle • Dispose of syringe in sharps container • Label bottles and send to lab Dr.T.V.Rao MD 19
  • 20. Method of Blood Collection • A minimum of 10 ml of blood is taken through venipuncture and injected into two or more "blood bottles" with specific media for aerobic and anaerobic organisms. • The blood is collected using clean technique. This requires that both the tops of the culture bottles and the venipuncture site of the patient are cleaned prior to collection with alcohol swabs containing 2% Chlorhexidine and 70% isopropyl alcohol. Dr.T.V.Rao MD 20
  • 21. The area of skin is cleaned with a disinfectant, or an alcohol swab. • Using sterile gloves, do not wipe away the surgical solution, touch the puncture site, or in any way compromise the sterile process. It is vital that the procedure is performed in as sterile a manner as possible as the persistent presence of skin commensals in blood cultures could indicate endocarditis but they are most often found as contaminants Dr.T.V.Rao MD 21
  • 22. The vein is anchored and the needle is inserted. Dr.T.V.Rao MD 22
  • 23. The vacutainer tube is depressed into the needle to begin drawing blood Dr.T.V.Rao MD 23
  • 24. Additional vacutainer tubes can be utilized. Determine what tests are ordered and what tubes will be necessary BEFORE you begin to draw blood, and determine the order of draw for the tubes. . Dr.T.V.Rao MD 24
  • 25. When the final tube is being drawn, release the tourniquet. Then remove the tube, and remove the needle. Dr.T.V.Rao MD 25
  • 26. After the needle is removed from the vein, apply firm pressure over the site to achieve haemostasis. Dr.T.V.Rao MD 26
  • 27. Apply a bandage to the area. Dr.T.V.Rao MD 27
  • 28. Preparation of Cap before Injecting Blood • Prep the rubber cap of the blood culture bottles with an alcohol pad in a circular motion. Allow the alcohol to dry. Dr.T.V.Rao MD 28
  • 29. Inject the Blood ….. • Inject the blood into the Selected Media • Gently rotate the bottles to mix the blood & the broth (do not shake vigorously). Dr.T.V.Rao MD 29
  • 30. Follow the universal precautions when disposing Needle • Dispose of needle in sharps container and dispose of other waste in proper container Dr.T.V.Rao MD 30
  • 31. Label the tubes, checking the requisition for the proper identification. Dr.T.V.Rao MD 31
  • 32. Give the all possible Medical Information • • • • • • • • Patient’s name • Hospital number (Patient ID) • Patient’s location (room and bed #) • Date and time of collection • Collector’s initials • Site of venipuncture • Or other information as per facility Include you Mobile Contact No – A vital information can be delivered any time Dr.T.V.Rao MD 32
  • 33. Document the Medical Records • Document the following in the medical record: • –Date & time specimen obtained • –Site of specimen collection Dr.T.V.Rao MD 33
  • 34. Frequency of Collection • Frequency of Collection – Depends if bacteremia is transient, intermediate or continuous – Number of cultures collected are usually inversely related to the type of bacteremia – Usually x3 from different body sites Dr.T.V.Rao MD 34
  • 35. Second Set • If 2 or more sets of blood cultures have been ordered, obtain the second set in the same manner as the first, making a new venipuncture at a different site. Dr.T.V.Rao MD 35
  • 36. Newer Blood Culture Methods • Newer Blood Culture Systems – Biphasic Broth-Slide System • Agar “paddles” attached to top of bottle • Closed system – Continuous Monitoring Blood Culture Systems • • • • BacTec – measures 14CO2 BacTec 9000 Series – measures CO2 ESP – measures consumption of gases BacT-Alert – measures change in pH Dr.T.V.Rao MD 36
  • 37. The Contaminated Blood Culture • If the skin is not adequately cleansed before drawing blood for culture, bacteria on the skin will be injected into the bottle, producing a false positive blood culture • It is difficult for the physician to determine whether the bacteria growing in the blood culture is a real pathogen causing bloodstream infection or whether bacteria on the skin have contaminated the culture. This can lead to excess use of antibiotics and prolongation of hospital stay. Dr.T.V.Rao MD 37
  • 38. Visit me for more articles of interest in Medicine and Health care please visit… Dr.T.V.Rao MD 38
  • 39. • The programme created by Dr.T.V.Rao MD as Technical Series for Microbiologists in the Developing World • Email • doctortvrao@gmail.com Dr.T.V.Rao MD 39