Pracactical Session of CVS Module

Blood Culture
 What is a blood culture?
A blood culture is a laboratory test in which blood, taken from the patient, is
inoculated into bottles containing culture media with the aim of growing
pathogenic bacteria or fungi for diagnostic purposes (to determine whether
infection-causing microorganisms are present in the patient’s bloodstream.
This is possible because the bloodstream is a sterile environment.
 What is the purpose?

Blood cultures are intended to:
 Confirm the presence of microorganisms in the bloodstream
 Identify the microbial etiology of the bloodstream infection
 Provide an organism for susceptibility testing and optimization of antimicrobial

therapy.
Blood culture bottles
Definitions:
 What is a bacteraemia?
The presence of viable bacteria in the bloodstream which may be

transient (e.g. following dental procedures), intermittent (e.g.
undrained abscesses), or continuous (e.g. endovascular infection).
 What is a septic shock?
A consequence of the host inflammatory response that can follow

infection associated with organ dysfunction, hypoperfusion or
hypotension.
 What is a fungaemia?
The presence of viable fungi in the bloodstream.

Indications for blood culture
1. Clinical features of sepsis including tachycardia, tachypnoea,

increased or sub-normal temperature and change in sensorium,
hypotension or prostration
2. Suspicion of infective endocarditis
3. Pyrexia of unknown origin
4. Unexplained leucocytosis or leucopenia
5. Systemic and localised infections including suspected meningitis,

osteomyelitis, septic arthritis, acute untreated bacterial pneumonia or
other possible bacterial infection
Growth medium
 Blood culture bottles contain a growth medium, which encourages
microorganisms to multiply, and an anticoagulant that prevents blood from
clotting.
 Sodium polyanethol sulfonate is the most commonly used anticoagulant because

it does not interfere with the growth of most organisms.
 The exact composition of the growth medium varies, but aerobic bottles use a
broth that is enriched with nutrients, such as brain-heart infusion or trypticase
soy broth, and anaerobic bottles typically contain a reducing agent such as
thioglycollate.
 The empty space in an anaerobic bottle is filled with a gas mixture that does not
contain oxygen.
Blood culture collection set
 Apron
 Clean non-sterile gloves
 Tourniquet
 Blood sampling device with blood culture bottle adapter (e.g.

winged blood collection set).
 Blood culture bottles (anaerobic and aerobic).
 Cleaning swabs (2% chlorhexidine in 70% isopropyl alcohol).
 Sterile gauze& plaster
 Laboratory forms, labels and transportation bag.

Blood culture collection set
The method
 Blood cultures should be collected: as soon as possible after the onset of clinical
symptoms; ideally, prior to the administration of antimicrobial therapy.
 Adult patient: a minimum of 10 cc. of blood is taken through venipuncture and

injected into two "blood bottles" with specific media for aerobic and
anaerobic organisms.
 Adequate volumes of blood improve detection of pathogenic organisms and

reduce time to detection.
 Pediatrics/Neonatal Patient: one aerobic bottle as ordered by the physician.

Recommended volume: 1 to 2 cc. of blood per bottle.
 In children, infection with anaerobic bacteria is uncommon, so a single aerobic

bottle may be collected to minimize the amount of blood required.
 Arterial blood culture provides no advantage over venous samples.
 The blood is collected using aseptic technique. This requires that both the tops
of the culture bottles and the venipuncture site of the patient are cleaned prior to
collection by swabbing with 70% isopropyl alcohol.
 To maximize the diagnostic yield of blood cultures, multiple sets of cultures

(each set consisting of aerobic and anaerobic vials) may be obtained.
 Two or more blood specimens should be collected using sterile technique at

separate sites, before administering antibiotics.
 Except in very unusual cases, no more than 3 sets of blood cultures should be
collected in one 24-hour period.
 A larger number of cultures may have to be collected from persons already
receiving antimicrobials, although, if clinical condition allows, stopping
antibiotics and re-culturing after 48 hours is preferred
 Obtaining multiple sets of cultures increases the probability of discovering
a pathogenic organism in the blood and reduces the probability of false positive
culture caused by skin contaminants .
 After inoculating the culture vials, advisably with new needles and not the ones
used for venipuncture, the vials are sent to the microbiology department.
 The bottles are entered into a blood culture machine (BACTEC blood culture
system), which incubates the specimens at body temperature. The blood culture
instrument reports positive blood cultures.
 However, negative growths do not exclude infection (culture –ve organisms).

Blood culture collection Automated BACTEC blood culture system
How many days of incubation are recommended?
 The current recommendation, and standard incubation period, for
routine blood cultures performed by continuous-monitoring blood
systems is five days, after which negative vials are removed.
 If a vial is positive, the blood is subcultured onto agar plates to

isolate the pathogenic organism for culture and susceptibility
testing.
 This culture and sensitivity process identifies the species of

bacteria and inform clinicians the appropriate antibiotics for
treatment.
Is it a contaminant or a true pathogen?
 Contamination of blood cultures during the collection process can produce
a significant level of false-positive results.
 A false positive is defined as growth of bacteria in the blood culture bottle
that were not present in the patient’s bloodstream, and were most likely
introduced during sample collection.
 Contamination can come from a number of sources: the patient’s skin, the
equipment used to take the sample, the hands of the person taking the blood
sample, or the environment.
 Contamination rates can be most effectively reduced by strict compliance with

hand hygiene rules and best practices for blood collection, particularly during the
stages of skin antisepsis, venipuncture and sample transfer to blood culture bottles.
CASE STUDY
A 47-year-old man who cut his thumb. The next morning, his thumb is sore and the
skin surrounding the cut is red. By the time, the thumb is swollen and throbbing, and
yellowish-white pus is oozing out of the injured area. He also notices two red
streaks going up the inside of his forearm. Just as he begins to think about getting
some medical attention for his thumb, the man experiences a shaking chill and
becomes queasy. His wife then drives him to the ER at the nearest hospital. Upon
their arrival at the ER, the man's temperature has reached 39.7ºC. He is flushed and
ill-appearing, with a pulse of 125 and a blood pressure of 100/60 mm Hg. (His
normal BP is 145/85 mm Hg.) There are no other remarkable findings on physical
examination.
What is your preliminary diagnosis?
What tests should you perform?
What are the most important causative agents overall?

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Blood Culture

 What is a blood culture?

This is possible because the bloodstream is a sterile environment.

 What is the purpose?

 Confirm the presence of microorganisms in the bloodstream

 Identify the microbial etiology of the bloodstream infection

 Provide an organism for susceptibility testing and optimization of antimicrobial

The presence of viable bacteria in the bloodstream which may be

 What is a septic shock?

A consequence of the host inflammatory response that can follow

The presence of viable fungi in the bloodstream.

1. Clinical features of sepsis including tachycardia, tachypnoea,

2. Suspicion of infective endocarditis

3. Pyrexia of unknown origin

4. Unexplained leucocytosis or leucopenia

5. Systemic and localised infections including suspected meningitis,

 Sodium polyanethol sulfonate is the most commonly used anticoagulant because

 Clean non-sterile gloves

 Blood sampling device with blood culture bottle adapter (e.g.

 Blood culture bottles (anaerobic and aerobic).

 Cleaning swabs (2% chlorhexidine in 70% isopropyl alcohol).

 Sterile gauze& plaster

 Laboratory forms, labels and transportation bag.

 Adult patient: a minimum of 10 cc. of blood is taken through venipuncture and

 Adequate volumes of blood improve detection of pathogenic organisms and

 Pediatrics/Neonatal Patient: one aerobic bottle as ordered by the physician.

 In children, infection with anaerobic bacteria is uncommon, so a single aerobic

 To maximize the diagnostic yield of blood cultures, multiple sets of cultures

 Two or more blood specimens should be collected using sterile technique at

 Obtaining multiple sets of cultures increases the probability of discovering

 However, negative growths do not exclude infection (culture –ve organisms).

 If a vial is positive, the blood is subcultured onto agar plates to

 This culture and sensitivity process identifies the species of

a significant level of false-positive results.

 A false positive is defined as growth of bacteria in the blood culture bottle

introduced during sample collection.

 Contamination rates can be most effectively reduced by strict compliance with

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