API (Analytical Profile Index) 20E Test
API (Analytical Profile Index) 20E is a biochemical panel for identification and
differentiation of members of the family Enterobacteriaceae.
Principle
The API range provides a standardized, miniaturized version of existing identification
techniques, which up until now were complicated to perform and difficult to read. In the
API 20E, the plastic strip holds twenty mini-test chambers containing dehydrated media
having chemically-defined compositions for each test. They usually detect enzymatic
activity, mostly related to fermentation of carbohydrate or catabolism of proteins or amino
acids by the inoculated organisms.
A bacterial suspension is used to rehydrate each of the wells and the strips are incubated.
During incubation, metabolism produces color changes that are either spontaneous or
revealed by the addition of reagents. All positive and negative test results are compiled to
obtain a profile number, which is then compared with profile numbers in a commercial
codebook (or online) to determine the identification of the bacterial species.
The Test Kit
The test kit enables the following tests:
1. ONPG: test for β-galactosidase enzyme by hydrolysis of the substrate o-
nitrophenyl-b-D-galactopyranoside
2. ADH: decarboxylation of the amino acid arginine by arginine dihydrolase
3. LDC: decarboxylation of the amino acid lysine by lysine decarboxylase
4. ODC: decarboxylation of the amino acid ornithine by ornithine
decarboxylase
5. CIT: utilization of citrate as only carbon source
6. H2S: production of hydrogen sulfide
7. URE: test for the enzyme urease
8. TDA (Tryptophan deaminase): detection of the enzyme tryptophan
deaminase: Reagent- Ferric Chloride.
9. IND: Indole Test-production of indole from tryptophan by the enzyme
tryptophanase . Reagent- Indole is detected by addition of Kovac’s reagent.
10. VP: the Voges-Proskauer test for the detection of acetoin (acetyl
methylcarbinol) produced by fermentation of glucose by bacteria utilizing
the butylene glycol pathway
11. GEL: test for the production of the enzyme gelatinase which liquefies gelatin
12. GLU: fermentation of glucose (hexose sugar)
13. MAN: fermentation of mannose (hexose sugar)
14. INO: fermentation of inositol (cyclic polyalcohol)
15. SOR: fermentation of sorbitol (alcohol sugar)
16. RHA: fermentation of rhamnose (methyl pentose sugar)
17. SAC: fermentation of sucrose (disaccharide)
18. MEL: fermentation of melibiose (disaccharide)
19. AMY: fermentation of amygdalin (glycoside)
20. ARA: fermentation of arabinose (pentose sugar)
Results
and
Interpretation
1. For some of the compartments, you can just read the change in color
straightway after 24 hours but for some, you have to put reagents before
reading.
2. Add the following reagents to these specific compartments
1. TDA: Put one drop of ferric chloride
2. IND: Put one drop of Kovacs reagent
3. VP: Put one drop of 40 % KOH (VP reagent 1) & One drop of VP
Reagent 2 (α-Naphthol) (you have to wait for 10 minutes before telling
it negative).
3. Get the API reading scale (color chart)
1. Mark each test as positive or negative on the lid of the tray
2. The wells are marked off into triplets by black triangles, for which
scores are allocated as follows:
4. Add up the scores for the positive wells only in each triplet. Supplementary
tests, e.g.: oxidase may also be included in the profile. The highest score
possible for a triplet is 7 (the sum of 1, 2 and 4) and the lowest is 0.
5. Numbering in API 20E Test Strip
1. The profile for this combination of reactions is therefore 7031645 (7 digit
code)
2. Identify the organism by using API catalog or apiweb
A partir de los siguientes resultados de las pruebas API, indique a que bacteria corresponde:
1. Pseudomonas aeruginosa 2206004
2. Salmonella typhi 4004540
3. Yersinia pestis 1004503
4. Enterobacter gergoviae 5315173
5. Vibrio cholerae 5347124
1. Pseudomonas aeruginosa 2206004
2. Salmonella typhi 4004540
3. Yersinia pestis 1004503
4. Enterobacter gergoviae 5315173
5. Vibrio cholerae 5347124
1. Pseudomonas aeruginosa 2206004
2. Salmonella typhi 4004540
3. Yersinia pestis 1004503
4. Enterobacter gergoviae 5315173
5. Vibrio cholerae 5347124
1. Pseudomonas aeruginosa 2206004
2. Salmonella typhi 4004540
3. Yersinia pestis 1004503
4. Enterobacter gergoviae 5315173
5. Vibrio cholerae 5347124
1. Pseudomonas aeruginosa 2206004
2. Salmonella typhi 4004540
3. Yersinia pestis 1004503
4. Enterobacter gergoviae 5315173
5. Vibrio cholerae 5347124