PRO DISC (BULK) K1532B- 50 tests . PRINCIPLE/DISCUSSION: Enzymatic hydrolysis of L-proline---ß-naphthylamide releases free-ß naphthylamine which is detected and shown by the color change after adding PEP reagent. This test is especially of use in screening for Clostridium difficile from CCFA. A negative test indicates the organism cannot be C. difficile. The test is also of value when used with other tests in the speciation of most anaerobes and other organisms such as enterics. Comprehensive charts for these organisms are available on request. MSDS The discs contain L-proline ß-naphthylamide: naphthylamides have been identified as possible carcinogens however when used as directed the discs present no hazard. PEP reagent contains hydrochloric acid and is mildly corrosive. PEP stains anything it contacts. Handle with care. Consult poison control center if ingested. MATERIAL REQUIRED: The tests require fresh 24 hour growth on media not available from KEY. Consult a suitable manual for recommended media for the specimen. The following items are required but not provided: Tube or slide Inoculating loop or stick Distilled water, pH 7.0-7.2 INSTRUCTIONS: (1) Place a disc onto a slide and moisten slightly. (Do not over saturate.) (2) Smear with a paste of the organism from a fresh pure 24 hour culture plate or slant. 3) Wait 2-5 minutes (at room temperature) then add 1 drop of PEP reagent. Wait at least 1 but not more than 2 minutes for color to develop. INTERPRETATION: The appearance of a dark pink to red color is positive. Indole reactions of organisms grown on any media containing tryptophane (e.g. blood agar) may interfere but will still produce predictable reactions. From such media, positive PRO reactions will range from dark pink or red (PRO+/indol-) to purple (PRO+/indole+). Green or turquoise (indole+), and yellow (indol-) are both PRO negative. STORAGE: Store discs and reagent tightly covered with desiccant in a dry place at 0-8 C. Discs may be used while cold. QUALITY CONTROL: Each lot should be tested with known positive and negative organisms. Some suggested strains are C. difficile ATCC# 9689 (positive) and C. perfringens ATCC #13124 (negative). Positive PRO and Indole may be shown by testing with C. sordelli. Dispose of all used material in a manner appropriate for hazardous ma-terial. REFERENCES (1) Manual of Clinical Microbiology, Fifth Edition, ASM, Washington,D.C. (2) Wadsworth Anaerobic Bacteriology Manual, 5th Edition, 1993,Glucosidase tests, page 152. 9811kb /HTML>FOR MSDS CLICK HERE