Blood Culture - Method

Method

A minimum of 10 ml of blood is taken through vein puncture and injected into two or more "blood bottles" with specific media for aerobic and anaerobic organisms. A common media used for anaerobes is thioglycollate broth.

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 with swabs 70% isopropyl alcohol.

To maximise the diagnostic yield of blood cultures multiple sets of cultures (each set consisting of aerobic & anaerobic vials filled with 3-10 mL) may be ordered by medical staff. A common protocol used in US hospitals includes the following:

• Set 1 = L. antecubital fossa at 0 minutes
• Set 2 = R. antecubital fossa at 30 minutes
• Set 3 = L. or R. antecubital fossa at 90 minutes

Ordering multiple sets of cultures increases the probability of discovering a pathogenic organism in the blood and reduces the probability of having a positive culture due to skin contaminants.

After inoculating the culture vials, advisably with new needles and not the ones used for venepuncture, they are sent to the clinical pathology microbiology department. Here the bottles are entered into a blood culture machine, which incubate the specimens at body temperature. The blood culture instrument reports positive blood cultures (cultures with bacteria present, thus indicating the patient is "bacteremic"). Most cultures are monitored for 5 days after which negative vials are removed.

If a vial is positive, a microbiologist will perform a Gram Stain on the blood for a rapid, general ID of the bacteria, which they will report to the attending physician of the bacteremic patient. The blood is also subcultured or "subbed" onto agar plates to isolate the pathogenic organism for culture and suceptibility testing, which takes up to 3 days. This culture & sensitivity (C&S) process identifies the species of bacteria. Antibiotic sensitivities are then assessed on the bacterial isolate to inform clinicians on appropriate antibiotics for treatment.

Some guidelines for infective endocarditis recommend taking up to 6 sets of blood for culture (around 60 ml).