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 and 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.

 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.

 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.

 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 .
 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.

 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.
 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).
False Negative Blood Culture Result
It may be due to following reasons:
i. Recent antibiotic therapy.

ii. Inadequate number of specimens: Repeated blood

cultures are necessary as bacteremia is intermittent.

iii. Infection with Coxiella burnetii or Chlamydia sp.

Blood cultures may be negative because Coxiella burnetii
and Chlamydia sp. cannot grow on cell free media.
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, or the hands of the person taking the blood
sample.

 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, the thumb is swollen and
throbbing, and yellowish-white pus is oozing out of the injured area. 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?