Effluent Culture & Gram Stain

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1. One set of Bactec Culture Bottles (1 aerobic and 1 anaerobic) and 1 Tall Yellow Top Tube are used for each Effluent Culture & Gram Stain ordered.
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2. Cleanse all sites for 30 seconds with a 70% Alcohol Prep Pad and another 30 seconds with iodine (e.g., Betadine) or chlorhexidine.
   • Note: Adequate disinfection minimizes contamination from skin and environmental organisms.
3. Remove flip-off dust caps from BACTEC Culture Bottles and disinfect rubber bottle tops with a 70% Alcohol Prep Pad, immediately before inoculating and allow to dry.
   • CAUTION: Do not use any form of iodine to disinfect the tops of the bottles. It may affect the integrity of the rubber top of the culture bottle, causing false positive results.
4. Push and hold Vacutainer holder over the top of the aerobic BACTEC Culture Bottle to puncture the septum. Allow the bottle to automatically fill until 8-10 mL collects.
   • CAUTION: Do not overfill or underfill BACTEC bottles, as this may reduce sensitivity and adversely affect the recovery of microorganisms.

5. Remove the aerobic BACTEC Culture Bottle from the holder.
6. Insert the top of the anaerobic BACTEC Culture Bottle into the holder (you may use the same holder). Allow the bottle to automatically fill until 8-10 mL collects.

7. Insert the stopper of the Yellow Top Tube into the holder and allow the tube to fill automatically until the flow ceases (one tube per test ordered).

8. Remove and discard the holder.
9. Use the barcode labels with the patient’s name and ID to identify each BACTEC Culture Bottle and the Yellow Top Tube.
   • CAUTION: Do not place the patient’s barcode label over the bottle’s barcode label.
10. Place in a transport bag.
11. Hold at room temperature.
12. Specimens are stable for 48 hours after collection.