1. Mycobacteriaceae – Part II Specimen Collection Created by: Michele L. Jurgensmeier MT(ASCP) Edited by: Kathy Talmon MT(ASCP)SM Voice over lecturer: Carol Larson MSEd, MT(ASCP) Clinical Laboratory Science Program

2. Mycobacteriaceae Specimen Collection Sterile disposable container Collect before antibiotic therapy Deliver promptly to laboratory Refrigerate if delay in delivery Swabs not acceptable in most laboratories

3. Mycobacteriaceae Specimen Types Respiratory - sputum, induced sputum, BAL Urine Gastric lavage Feces (not accepted in most labs) Wound drainage Blood Body fluids Tissues

4. Mycobacteriaceae Respiratory Specimens Sputum, induced sputum and BAL Irregular and intermittent release of mycobacteria into bronchial lumen –Recommend minimum of 3 to 5 early morning specimens on consecutive days 10 mL or less per 50 mL tube NO pooled specimens

5. Mycobacteriaceae Urine Specimens 3 to 5 early morning midstream or catheterized specimens Entire volume or minimum 15 mls Centrifuge to concentrate specimen

6. Mycobacteriaceae Gastric Lavage Obtained when unable to obtain sputum samples Preferred specimen for children Must neutralize acids in specimen

7. Mycobacteriaceae Feces NOT acceptable for culture Some institutions do accept this specimen –Immunocompromized patients (AIDS) Risk for disseminated mycobacterial disease

8. Mycobacteriaceae Aspirated Wound Drainages Fluid collected Swabs not acceptable

9. Mycobacteriaceae Blood Cultures Lysis centrifugation –Release intracellular mycobacteria –Concentrate organisms if present Or inoculate directly to broth media Direct inoculation to solid media should not be done

10. Mycobacteriaceae Sterile Body Fluids Sterility strictly maintained Centrifuge and plate sediment CSF –Large volumes needed –Difficult to diagnosis tuberculous meningitis

11. Mycobacteriaceae Tissues Sterility strictly maintained Saline added to avoid dehydration In the lab, tissue is ground to release organisms

12. Mycobacteriaceae In review… Specimen collection –Mycobacteria isolated from variety of specimens Most common respiratory –Sterile technique in collecting body fluids and tissue –Concentrate specimens since mycobacteria may be present in small numbers

13. Mycobacteriaceae References: 1.Mahon, C.R. & Manuselis, G., Textbook of Diagnostic Microbiology, 2nd Ed., W.B. Saunders, 2000 2.Forbes, B.A., Sahm, D. F., & Weissfeld, A.S., Bailey and Scott’s Diagnostic Microbiology, 11th Ed., Mosby, 2002. 3.Koneman, Color Atlas and Textbook of Diagnostic Microbiology, 5 th Ed., J.B. Lippincott Co., 1997 4.Murray PK, et al., Manual of Clinical Microbiology, 7th ed., ASM Publishing, Washington DC, 1999.