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API
API
This API-20E test strip (from bioMerieux, Inc.) is used to identify the enteric gram negative
rods (although API makes a variety of other test strips for yeast, Staph, anaerobes, etc.) 20
separate test compartments are on the strip, all dehydrated. A bacterial suspension is used to
rehydrate each of the wells. Some of the wells will have color changes due to pH differences:
others produce end products that have to be identified with reagents. A profile number is
determined from the sequence of + and - test results, then looked up in a codebook having a
correlation between numbers and bacterial species.
OBJECTIVE:
Learn how to perform and interpret the miniaturized, multi-test technique for bacterial
identification.
MATERIALS NEEDED:
PROCEDURE:
1. Inoculate a large colony (2-3mm diameter)of the bacterium (pure culture) into the
0.85% NaCl solution, making sure that the suspension is
homogenous and without clumps of floating bacteria.
2. Use a McFarland barium sulfate standard #3 to quantitate the
suspension and to produce a standard inoculums size..
1. The bottom of the incubation chamber has small indented wells in the bottom: fill it with
water just enough to fill these indentations.
2. Place the strip into this bottom. There should not be so much water that it slops onto
the API strip.
3. Place the top of the incubation chamber over the bottom, and label it.
4. Place the strip at 37o C for 18-24 hours.
INTERPRETATION:
QUESTIONS:
3. What are the advantages of this test (compared to regular biochemical tube media)?