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Mycobacteriaceae – Part IV Microscopic Examination

Mycobacteriaceae – Part IV Microscopic Examination. 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. Microscopic Examination.

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Mycobacteriaceae – Part IV Microscopic Examination

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  1. Mycobacteriaceae – Part IVMicroscopic Examination 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. Microscopic Examination • Direct specimen detection important due to slow growth rate of organisms • Purposes • Determine acid fast characteristic • Detect new cases • Monitor progress of disease / treatment • Criteria for patient release from hospital

  3. Microscopic Examination • Fluorochrome stains (Auramine O) • Primary stain: fluorescent dyes • Counter stain: potassium permanganate • AFB fluoresce yellow/gold against a black background Enlarged view

  4. Microscopic Examination • Fluorochrome stains (Auramine O) • Screening procedure • Advantages • More sensitive than carbolfuchsin stains • Examine on lower magnification • View more fields in shorter time • Disadvantages • Rapidly growing mycobacteria do not stain • Less specific

  5. Microscopic Examination • Carbolfuschin stains (Ziehl-Neelsen / Kinyoun) • Primary stain: carbolfuschin • Counter stain: methylene blue • AFB appear red and background material blue Photo courtesy of CDC public image library Content provider Dr. George P. Kubica

  6. Microscopic Examination • Carbolfuschin stains (Ziehl-Neelsen / Kinyoun) • Advantages • More specific than fluorescent stains • Stains all mycobacteria • Disadvantages • Longer examination time • Less sensitive

  7. Microscopic Examination • Interpretation of smears • Examine 300 oil immersion fields • Observe slides for morphology consistent with mycobacteria • Other organisms may stain partially acid fast • Quantify organisms • Follow disease state

  8. Microscopic Examination –Correlation With Culture Results • Extensive disease • Good correlation • Minimal or less-advanced disease • Less correlation • Antibiotic treatment • Unsatisfactory laboratory procedures

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

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