Lab Test Procedure : Blood Cultures

Document Control Version 4

Special Notes To prevent specimen contamination by microorganisms, use sterile supplies and aseptic technique to collect specimens. Obtain cultures prior to starting antibiotic therapy. Notify the physician if unable to obtain cultures.

Venipuncture needle
Syringe for specimen
Chlorhexidine Prep
70% isopropyl acohol pad
Adhesive bandage

Blood Culture Media Always consider patient's weight when choosing appropriate bottles (See Recommended blood volumes in chart below)

Collection Procedure
1. Clean venipuncture site with 70% isopropyl alcohol (alcohol pad), allow to air dry, and follow with Chlorhexidine Prep using back and forth friction scrub for 30 seconds. Allow it to dry for 30 seconds.
2. Do not retouch site; if you must repalpate for vein, the area must be recleaned using above procedure. Perform venipuncture.
3. When required amount of blood is received, remove needle from skin. NOTE: When collecting blood through an intravenous line, it is not necessary to discard the initial volume of blood.
4. Hold pressure to site with gauze and apply adhesive bandage when bleeding stops.
5. Remove venipuncture needle from syringe and attach blunt filled needle to transfer device.
6. Remove tops of blood culture bottles and clean with 70% isopropyl alcohol and allow to dry.
7. Inject blood specimens into bottles, place specimen into the anaerobic (orange) bottle, then the aerobic ( green depending on patient's weight)bottle. If you only have enough blood for one bottle, place the specimen in the aerobic (green) bottle. Be careful, the bottles have vacuum and will aspirate all of the blood from the syringe unless you control the flow with your thumb and index finger. Optimal blood amounts are necessary or you will have decreased sensitivity which may delay time detection of a true positive blood culture.
8. Label bottle. Indicate whether line draw or peripheral.
9. Discard needle and syringe into sharps container.
10. Send specimen promptly to CCL. If the specimen is coming from the floors, send the audit trail with specimen that includes initials of person drawing the specimen, date, time and venipuncture site.

Bowden, V.R., Smith Greenberg, C.(2008). Pediatric Nursing Procedures, 2nd Ed. Philadelphia: Wolters Kluwer/Lippincott Williams & Wilkins.
Isenbery, H.D. (2004). Clinical Microbiology Procedures Handbook, Volume 2. Washington, D.C.: The ASM Press.

Blood culture recommended volume based on patient weight


 Patient weight in Kg

Blood volume to collect

Media type and  inoculation volume




(green FA plus)


(orange FN Plus)

1 to 3  (<6.6 lbs.)


1 ml


3.1 to 6 (6.6-13 lbs)


2 ml


6.1 to 9 (13-20 lbs)

4 ml

4 ml


9.1 to 12 (20-26 lbs)


4 ml

2 ml

12.1 to 20 (26-44 lbs)


4 ml

4 ml

20.1 to 25 (44-55 lbs)


5 ml

5 ml

25.1 to 40 (55-88 lbs)



5 ml

> 40 (>88 lbs)



10 ml

(revised 3-25-15)



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