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Standard Operating Procedure References Applicable to Detail Standard Operating Procedure Subject KOH Prep for Fungal Elements Index Number Lab-8649 Section Laboratory Subsection Regional/Affiliates Category Departmental Contact Rachel Blum Last Revised 9/17/2019 References Required document for Laboratory Accreditation by the College of American Pathologists (CAP), Centers for Medicare and Medicaid Services (CMS) and/or COLA. Applicable To Employees of the Gundersen Lutheran Clinical laboratories, Gundersen Tri-County Hospital laboratories, Gundersen St. Joseph’s Hospital laboratories, Gundersen Boscobel Hospital and Clinics laboratories, Gundersen Palmer Lutheran Hospital and Clinics laboratories and Gundersen Moundview Hospital and Clinic laboratories. Detail PRINCIPLE: The KOH mount is used to aid in detecting fungal elements in specimens containing keratinous material, such as skin, nails, or hair. The KOH dissolves the background keratin, unmasking the fungal elements to make them more apparent. Dissolving is improved by gentle heating, using the incubator or other source of generated heat. Open flames are discouraged because of the risk of fire. CLINICAL SIGNIFICANCE: Dermatophytosis is a clinical condition caused by fungal infection of the skin, hair and nails. The fungi that cause parasitic infection (dermatophytes) feed on keratin, the material found in the outer layer of skin, hair, and nails. These fungi thrive on skin that is warm and moist, but may also survive directly on the outsides of hair shafts or in their interiors. SPECIMEN: Skin scrapings, nail scrapings or hair shafts; Specimen will be collected by a physician scraping particles from the affected area onto a clean slide. Specimen slide is labeled with the patient’s name, date and medical record number and delivered to the lab. REAGENTS/MATERIALS: 1. 10% potassium hydroxide (KOH) (This reagent has a one year expiration date when stored at room temperature.) 2. Clean slide and coverslip EQUIPMENT/INSTRUMENTATION: 1. Heat source (incubator or slide warmer) 2. Microscope Page 1 of 3 Standard Operating Procedure CALIBRATION: N/A QUALITY CONTROL: No commercial controls are available. External Quality Control: Affiliate hospitals: external QC will auto order each day. If no patients are run, delete QC. If another KOH test is ordered after the first one of the day, QC will need to be manually ordered. Regional Clinic Labs: order external QC as needed with each patient test. Result with known QC pictures. Report External QC as Positive KOH QC picture or Negative KOH QC picture Quality Control results will be reviewed monthly by the Technical Leader or designee. Proficiency testing performed as required. Daily Quality Control POSITIVE QC CONTROL SLIDES NEGATIVE QC CONTROL SLIDES Positive KOH Negative KOH Implementation PROCEDURE: 1. Suspend fragments of skin scales in either one or two drops of 10% KOH. 2. Apply coverslip and heat on slide warmer. Allow to stand 5-10 minutes. 3. Examine preparation completely for hyphae segments, spores, budding yeast, mycotic granules, or spherules. a. Hyphae are tubular, usually branching and septate, cells which make up the vegetative portion of mycelium of fungi. b. Spores are the reproductive forms of fungi. c. Budding yeast and the round, thicker walled spherules are also reproductive forms of fungi. 4. Upon completion of the examination, discard slide into sharps biohazard container. 5. Report out as: “No fungal elements present” or “Fungal elements present” in the LIS. Page 2 of 3 Standard Operating Procedure CALCULATIONS: N/A INTERPRETATION: N/A LIMITATIONS: N/A REVIEW AND CHANGES: This document should be reviewed optimally on an annual basis, with 2 years as the maximum review date. Review will be done by the Technical Leader, Supervisor, Manager, Medical Director, or designated person. Changes require retyping document or form and review by the Medical Director. REFERENCES: 1. Medical Mycology, Emmons, Binford & Utz, 1964. 2. Bailey & Scott’s. Diagnostic Microbiology, 12th edition. Ellen Jo Barron and Sydney M. Finegold, 1990. Page 3 of 3 .
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