Blood Culture Collection

ARM DECONTAMINATION

Select a venipuncture site that is free of infection or other skin disorders. Decontamination of the site is required to prevent infection of the patient or contamination of the blood culture devices.

Procedure:

• Use the ChloraPrep One-Step Frepp Applicator containing 2% chlorhexidime gluconate and 70% isopropyl alcohol.
  • Note: Do not use ChloraPrep on patients <=2 months old.  Use 70% isopropyl alcohol as a disinfectant to the venipuncture site.
• Remove the ChloraPrep from the sterile packaging.
• Break the ampule by pressing the plastic wings together.
• Allow the solution to saturate the sponge pad.
• Use a scrubbing motion to apply disinfectant to the venipuncture site.
• Allow the disinfectant to dry for 30 seconds prior to performing the venipuncture.

FREQUENCY OF DRAWING

In some infections, bacteremia is continuous and several blood cultures can be taken at the same time but preferably from different body sites. With other conditions, however, bacteremia is intermittent. There will be times when bacteria can be recovered from the blood interspersed with times when the blood is sterile. To cover both the possibilities of intermittent or continuous bacteremia, the general rule is to draw three blood cultures, one-half hour apart.

For almost all clinical situations, the laboratory recommendation is to collect from each patient, a minimum of two blood cultures and a maximum of three blood cultures, one-half hour apart. Alternatively, multiple blood cultures can be collected at the same time provided they are collected from different venipuncture sites.

Drawing Exceptions:

• If the physician wants to begin antimicrobial therapy immediately, draw two or three blood cultures at the same time, using two different sites if possible.
• If antimicrobial agents have already been administered, 4 to 6 separate blood cultures may be necessary to recover a pathogen.
• If endocarditis is suspected, try to draw three blood cultures at no less than hourly intervals within a 24 hour time period prior to initiation of antimicrobial therapy if possible.
• If the physician does not want the cultures all drawn at one time, but does want them at time intervals of less than one-half hour, the laboratory policy is for the physician (or someone delegated by him) to draw the blood for cultures and have the bottles sent down to the laboratory. The lab will supply the bottles.
• If the patient is at home or at a nursing home, the recommendation is that two or three blood cultures should be collected at the same time but from different sites.

BLOOD COLLECTION & INOCULATION

• For adult routine blood cultures collect about 20mL blood using a sterile needle and syringe, or a blood collection set.
• Wipe the tops of the blood culture devices with isopropyl alcohol, to disinfect. Always inject the blood into the aerobic device first then into the anaerobic bottle. Any air in the blood culture set will be inoculated into the aerobic system only.
• Enter the following information on the device labels: patient’s name, collection time, and the initials of the phlebotomist. Do not cover the bottle bar code label.
• For premature infants weighing more than 1 kg (2.2 lbs.) to 2 kg (4.4 lbs.) collect 4.5 mL of blood.  Inject 4.5 mL into the aerobic bottle BacTec Peds Plus/F.
• For infants weighing more than 2 kg (4.4 lbs.) to 12.7 kg (28 lbs.) collect 5.0 mL of blood. Inject 5.0 mL into the aerobic bottle BacTec Peds Plus/F.
• For children and adults weighing between 12.7 kg (28 lbs.) and 25 kg (55 lbs.) collect 10.0 mL of blood per set. Inject 5.0 mL each into an adult BacTec Plus + aerobic bottle and 5.0 mL into a BacTec Lytic/10 anaerobic bottle.
• For children and adults weighing >25 kg (55 lbs.) and 36.3 kg (80 lbs.) collect 21.5 mL of blood per set. Inject 10 mL into a BacTec Plus + aerobic bottle and 10 mL into a BacTec Lytic/10 anaerobic bottle. 
• For children and adults weighing more than 36.3 kg (80 lbs.) collect 20 mL of blood.  Inject 10 mL into a BacTec Plus + aerobic bottle and 10 mL into a BacTec Lytic/10 anaerobic bottle.  Preferably in one-half hour, collect an additional 20 mL of blood to be inoculated as above.

Note:  If a Catheter Related Sepsis (Quantitative), fungal, or mycobacterial blood culture is ordered, draw a 10mL Isolator tube for each of these procedures in addition to the routine set of bottles.

TRANSPORTATION TO THE LAB

The inoculated bottles must be brought to the lab and placed in the BacTec Blood Culture System as soon as possible to minimize the detection time of positive cultures. When bottles are held at room temperature for a prolonged time, any bacteria in them will revert from the log phase of growth to the lag phase, slowing the recovery by as much as one day or more.

ROUTINE BLOOD CULTURE COLLECTION CHART

Age	Weight	Interval time of collection	Number of Cultures	Recommended volume of blood per tube or bottle
Premature Newborns	≤ 1 kilogram (2.2 lbs)	N/A	1	2 mL total volume 2 mL in aerobic bottle BACTEC PEDS PLUS / F  (pink)
Premature Newborns	>1-2 kilograms (>2.2-4.4 lbs)	N/A	1	4.5 mL total volume 4.5 mL in aerobic bottle BACTEC PEDS PLUS / F  (pink)
Full term Infants to 2 years	>2-12.7 kilograms (>4.4-28 lbs)	N/A	1	5 mL total volume 5 mL in aerobic bottle BACTEC PEDS PLUS / F  (pink)
Children >2 to 5 years	>12.7- 25.0 kilograms (>28-55 lbs)	N/A	1	10 mL total volume 5 mL in BACTEC aerobic bottle (blue) 5mL in BACTEC anaerobic bottle (purple)
Children >5 to 10 years	>25.0 – 36.3 kilograms (>55-80 lbs)	N/A	1	20 mL total volume 10 mL in BACTEC aerobic bottle (blue) 10 mL in BACTEC anaerobic bottle (purple)
Children and adults over 80 lbs	>36.3 kilograms (>80 lbs)	Collect 2 sets of blood cultures at the same time from 2 different venipuncture sites or from the same site 5 minutes to ½ hour apart	First 2 cultures           Additional cultures	40-60 mL total volume   Collect 20 mL per set: 10 mL for each BACTEC aerobic (blue) 10 mL for each BACTEC anaerobic (purple)   Collect 20 mL per set: 10 mL for each BACTEC aerobic (blue) 10 mL for each BACTEC anaerobic (purple)

Special order blood cultures for children and adults over 36.3 kilograms (>80 lbs):

1. Vascular access device blood cultures:  Nurse to draw bottles as listed above.
2. Catheter related sepsis blood cultures:  Nurse to draw one 10 mL isolator from the device.  Lab to draw 10 mL Isolator from the venous site.  Both are listed under one accession number.  See separate procedures for complete instructions.
3. Autologous blood cultures:  Collect blood culture bottles as described above.  Record unit number.
4. Fungal cultures:  Collect one 10 mL Isolator per set.
5. Mycobacteria cultures:  Collect one 10 mL Isolator per set. 

Container	Recommended Volume	Minimum Volume
Adult Isolator	10 mL	6 mL
BACTEC PEDS PLUS / F bottle	1-5 mL	0.5 mL
BACTEC aerobic and anaerobic bottles	10 mL	3 mL

NOTES:

1. BACTEC PEDS PLUS (pink) and aerobic bottle (blue) must be collected before the BACTEC anaerobic bottle (purple)./p>
2. Blood cultures must be collected as soon as possible and no later than 30 minutes after they were ordered by the physician.
3. Do NOT use ChloroPrep® on infants ≤2 months of age.  Use alcohol only for disinfection of venipuncture site in this age group.