Martin Health System Stuart, Florida Laboratory Services Microbiology Specimen Collection The recovery of pathogenic organisms responsible for an infectious process is dependent on proper collection and transportation of a specimen. An improperly collected or transported specimen may lead to failure to isolate the causative organism(s) of an infectious disease or may result in the recovery and subsequent treatment of contaminating organisms. Improper handling of specimens may also lead to accidental exposure to infectious material. GENERAL GUIDELINES FOR COLLECTION: 1. Follow universal precaution guidelines, treating all specimens as potentially hazardous. 2. Whenever possible, collect specimen before antibiotics are administered. 3. Collect the specimen from the actual site of infection, avoiding contamination from adjacent tissues or secretions. 4. Collect the specimen at optimal time i.e. Early morning sputum for AFB First voided specimen for urine culture Specimens ordered x 3 should be 24 hours apart (exception: Blood cultures which are collected as indicated by the physician) 5. Collect sufficient quantity of material for tests requested. 6. Use appropriate collection and transport container. Refer to the Microbiology Quick Reference Chart or to the specific source guidelines. Containers should always be tightly sealed and leak-proof 7. Label the specimen with the Patient’s First and Last Name Date of Birth Collection Date and Time Collector’s First and Last Name Source 8. Submit specimen in the sealed portion of a Biohazard specimen bag. 9. Place signed laboratory order in the outside pocket of the specimen bag. 10. Include any pertinent information i.e. recent travel or relocation, previous antibiotic therapy, unusual suspected organisms, method or environment of wound infliction 11. Transport at the proper temperature, i.e. room temp, 2-8c and deliver to the Laboratory as quickly as possible. Organism Identification and Sensitivity will be performed on all significant isolates. CLM.20 1 Specific Collection and Transportation Guidelines: 1. Abscess Culture: Please refer to Wound/Abscess 2. Aerobic Culture: Please refer to Source 3. Anaerobic Culture: *Must be ordered with an Aerobic Culture Container: Anaerobic Specimen Collector (one per site) *Do not use expired tubes or if the indicator (E) is pink Transport: Room Temperature Specimen: Wound, Tissue, or Abscess The best specimen for anaerobic culture is obtained using a needle and syringe. When a swab must be used to collect a specimen, an Anaerobic Specimen Collector should be used. Special care must be taken to sample the active site of a deep wound infection. Unacceptable Specimens: Superficial Wound sites Throat or Nasopharyngeal Swabs Gingival or Mouth Swabs Expectorated or Induced Sputum Voided or Catheterized Urine Vaginal or Cervical Swabs Specimen Collection Procedure: *Do not remove stopper during collection. a. Collection by Swab: 1) Peel apart package (G) and remove specimen collector. 2) Remove plunger with sterile swab attached (B). 3) Decontaminate skin surface using 70-95% alcohol. Obtain specimen. 4) Replace swab through holes and into inner tube (C). 5) Hold tube at a 45° angle. Press down on disc portion of plastic plunger (B) with continuous gentle force until plastic disc rests against top of rubber stopper (A) forcing the inner tube (C) into outer tube (F). CLM.20 2 Note: The hole will be plugged by the plastic plunger and the small inner tube will be detached from the stopper. 6) Hold tube at approximately a 10 - 30° angle and rotate with a swirling motion. This is to facilitate mixing of air in inner tube with the hydrogen atmosphere in outer tube. 7) Transport to laboratory. For collectors containing liquid specimens, maintain in an upright position. b. Collection of Liquid or Purulent Specimens: 1) Decontaminate the skin surface using 70-95% alcohol. 2) Collect specimen with sterile syringe and needle or a flexible plastic catheter. 3) Air trapped in syringe should be expelled by holding syringe and needle upright. Expel air at tip of syringe into alcohol saturated sponge. 4) Peel apart package (G) and remove specimen collector. 5) Remove swab plunger unit (B) and expel liquid material into inner tube (C). (Tissue: Aseptically cut tissue into small pieces and place in inner tube) 6) Replace swab through holes into inner tube (C). 7) Hold tube at a 45° angle. Press down on disc portion of plastic plunger (B) with continuous gentle force until plastic disc rests against the rubber stopper (A) forcing the inner tube (C) into the outer tube (F). Note: The hole will be plugged by the plastic plunger and the small inner tube will be detached from the stopper. 8) Hold tube at approximately a 10 - 30° angle and rotate with a swirling motion. This is to facilitate mixing of air in inner tube with the hydrogen atmosphere in outer tube. 9) Transport to laboratory. For collectors containing liquid specimens, maintain in an upright position. 4. Beta Strep Culture: Throat - Refer to collection procedure for Throat Culture. Specify Beta Strep Screen Prenatal Screen - please collect vaginal and rectal specimen for Group B Strep 5. Biopsy/Tissue/Aspirate: Container: Sterile Specimen Container Syringe with safety cap – No needles! CLM.20 3 Anaerobic Transport vial Universal Transport Media (UTM) Specimen: Abscess, Tissue, Wound A few drops of sterile non-bacteriostatic saline or water may be added to prevent desiccation. If an anaerobic culture is desired, transfer a portion of the biopsy tissue to an Anaerobic transport vial. For Viral Culture, place a portion of tissue in Universal Transport Media. Do not submit tissue for culture in formalin. If surgical pathology is also desired, please submit a separate biopsy/tissue in formalin with a Pathology Requisition. Submit specimens in separate biohazard bags. If only one specimen is obtained and both culture and pathology are desired, please contact the Laboratory at 223-5945, Ext. 6928 (Microbiology) or Ext. 3146 (Histology) for further instructions. Preferred specimens for sinus tract: Aspiration material obtained by needle or catheterization Curetting from lining of sinus tract. Swabs specimens of sinus tract are acceptable only if the above cannot be obtained. (Swabs of sinus tract may not accurately reflect underlying disease process) Transport: Room Temperature 2 - 8°C (UTM) Specimen Collection Procedure: 1) Aseptically transfer tissue to a sterile container. 2) Include material from both the center and edges of lesion. 6. Blood Culture: (See Blood Culture Procedure, LSCM.11) 7. Body Fluid Culture: Excludes CSF and Urine Container: Sterile Specimen Container Sterile conical tube Sterile Syringe w/ Safety Cap (No needles) Aerobic Blood Culture bottle – peritoneal fluid only (10 mL) Plain Red Vacutainer tube (No gel) Anaerobic Specimen Collector Universal Transport Media (UTM) Volume: 5 – 10 mL per test (Minimum of 1 mL) CLM.20 4 Transport: Room Temperature 2 - 8°C (UTM) Specimen Collection Procedure: 1) Vigorously cleanse aspirate site for one minute with Chlorascrub skin disinfectant. 2) Allow Chlorascrub to air dry. 3) Collect the fluid using sterile needle and syringe or evacuated bottles. (A small amount of sterile heparin may be added to prevent clotting) 4) Aliquot specimen into the appropriate containers. Note: 10 mL of peritoneal fluid may be added to an aerobic blood culture bottle. (Peritoneal is the only fluid that should be cultured in blood culture media) 8. Bone Marrow Culture: Container: Standard Aerobic Blood Culture bottle (Blue) MycoF/Lytic Blood Culture bottle (White) EDTA tube Dark Green tube Volume: 3 -5 ml for Blood Culture bottles 4 ml for EDTA and Dark Green tubes Transport: Room Temperature Specimen Collection Procedure: 1) If using blood culture bottles, remove the protective shield from the bottles. 2) Cleanse the rubber stopper with Alcohol. 3) Physicians should wear gowns, masks and gloves during specimen collection. 4) Open ChloraScrub one-step applicator. 5) Prep site with ChloraScrub. Do not use ChloraScrub for children less than 2 months. Use alcohol, followed by betadine. Pinch the wings of the applicator to break the ampule and release the antiseptic. Press swab against the treatment area using repeated back and forth strokes for approximately 30 seconds to completely wet the treatment area. Allow to air dry for approximately 30 seconds. Do not blot, blow on or wipe the area. 6) Drape the surrounding skin with sterile drape. 7) Aspirate the marrow percutaneously using a sterile needle and syringe. 8) Immediately transfer the specimen to the appropriate containers. CLM.20 5 9. Bordetella pertussis Culture and DFA: Specimen: Nasopharyngeal Container: Regan/Lowe Bordetella Transport: Room Temperature Specimen Collection Procedure: Refer to Nose/Nasopharyngeal Note: DFA stain, in addition to culture, is suggested for diagnosis of Bordetella pertussis. 10. Catheter Tip Culture: Container: Sterile Container Specimen: 2 inch segment of catheter tip Transport: Room Temperature Specimen Collection Procedure: 1) Disinfect the intravenous catheter insertion site with Chlorascrub. 2) Withdraw catheter and snip off catheter tip, above the site of skin contact with sterile scissors. 3) To prevent desiccation, place sterile gauze moistened with sterile water on the bottom of the transport cup. Place catheter tip on top of the moistened gauze. Unacceptable Specimen: Urinary Foley Catheter Tips 11. Cerebrospinal Fluid Culture: Specimen: CSF Container: