API (Analytical Profile Index) 20E presented is a biochemical panel for identification and differentiation of members of the family. Changes to thesaurus. Changes to database old new. Taxons. Notes. Identification. Additional tests. API® 20 E v v X. X. X. X. RapiD 20 E™ v v X. API® 20 Microbial Identification Kits, bioMerieux. Supplier: Quantity. API® 20 E Microbial Identification Kit, , , , Pack of 25, Retrieving.
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Organism identification is based on pH change and substrate utilisations as established by published reference methodologies 4,6,8,9. Refer to the Table of Reactions below for the substrates contained in each wpi, the specific reaction principle, and colour changes. Each strip consists of 12 different biochemical substrates. The 12A strip may be used alone for identification of oxidase-negative, nitrate-positive glucose fermenters comprising 15 genera and may be useful for screening pathogenic Enterobacteriaceae from enteric and urine specimens or identification of other common isolates.
The 12B strip can be used in conjunction with the 12A strip for the identification of oxidase-positive, nitrate-negative, and glucose-nonfermenters MGNB as well galeriee the Enterobacteriaceae. The 12 substrates contained in the 12A strips are available in a solid microplate format, galfrie to as 12E.
The 12B strips can be used alongside the 12E, but in a separate tray. The 24E solid microplate format contains the 24 substrates contained in the combination of both the 12A and 12B strips. Galreie and precautions 1. These strips are intended for in vitro halerie only; for use by qualified laboratory personnel using aseptic techniques and established precautions against microbiological hazards.
Used materials should be autoclaved, incinerated, or immersed in germicide before disposal. Once the foil pouch has been opened, unused strips must be placed back in the foil pouch, and the foil pouch taped closed.
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Strips stored in this manner must be used within 7 days. Kit presentation Each kit contains the following: Materials Required but not provided The following materials may be required but are not provided:.
These can be purchased individually or as a balerie by ordering product code MB Set up proceedure Isolation An hour pure culture of the organism to be identified must be obtained.
Qualified Personnel should collect specimens according to standards routinely required for specimen handling Before use, perform an oxidase test on the organism to be identified.
Identification of Pseudomonas aeruginosa with the API-20E system.
Refer to the procedure chart for a condensed version of the following procedures. Preparation of inoculum Pick isolated colonies from an hour culture and emulsify in 2. Mix thoroughly to prepare a homogeneous suspension. If the organism has been grown on a selective medium and the colony is small or inhibited, it may be necessary to emulsify the colony in 5. Using a sterile Pasteur pipette, transfer one drop of the peptone water culture into the appropriate volume see Procedure Chart of sterile saline solution 0.
Inoculation The wells of individual substrate sets can be exposed by cutting the end tag of the sealing strip and slowly peeling it back. When Actinobacillus or Pasteurella sp. Using a sterile pipette or dropper bottle, overlay the substrates underlined on the holding tray with sterile mineral oil, i.
Well 8 for 12B and 20 for 24E is not overlayed with oil for oxidase- positive, miscellaneous Gram-negative bacilli. Incubation Reseal the inoculated rows with the adhesive seal and write the specimen identification number on the end tag galfrie a marker pen.
To determine the purity of the inoculum, xpi is advisable to inoculate a solid non-selective medium with the test suspension to act as a culture purity check. Reading the test strip 1. The 12A 12E strip should be read at hours. Aglerie systems should be read after 48 hours for the identification of Miscellaneous Gram-negative bacilli. Remove the strips or tray from the incubator, peel back the sealing tape. Record all positive results.
The reactions are evaluated as positive or negative by comparing them with the colour chart. Record the results under the appropriate heading on the report form. For aid in interpreting reactions, refer to the Table of Reactions. Well 8 Indole production – add 2 drops of Indole Kovacs reagent. Evaluate within 2 minutes of the addition of the reagent. Evaluate 15 to 30 minutes after the addition of reagents. Test can be evaluated immediately after the addition of the reagent.
Hydrolysis of gelatin is indicated by dispersal of the black particles throughout the well. The arginine reaction well 12 for 12B and well 24 of 24E is interpreted differently at 24 hours and 48 hours incubation. Yellow-green – Negative Blue – Positive. One drop of Nitrate reagent A and 1 drop of Nitrate reagent B is added to the well. Production of a red colour within a few minutes of the addition of the reagent indicates that nitrate reduction to nitrite NO 2 has occurred.
A small amount of zinc powder should be added to those wells which exhibit a yellow colour after the addition of the nitrate reagents. This will determine whether nitrate has been reduced completely to nitrogen gas N2.
The results should be interpreted as follows:. All organisms belonging to the family Enterobacteriaceae reduce nitrates to nitrites and give a positive reaction.
Bromothymol blue indicator changes from blue to yellow when the carbohydrate is utilised to form acid.
Indole is formed from metabolism of tryptophan. Indole Kovacs reagent forms a pink-red complex with indole. Tryptophan deaminase forms indolepyruvic acid from tryptophan which produces a brown colour in the presence of ferric ions.
Indole positive organisms may produce a brown colour. This is a negative reaction. Bromothymol blue indicator changes from blue to yellow when the carbohydrate is fermented.
Argine dihydrolase converts arginine into ornithine, ammonia and carbon dioxide. This causes a pH rise as indicated by bromothymol blue. Green reactions occurring at 48 hours should be interpreted as negative. Each group of three reactions produces a single digit of the code. Using the results obtained, the indices of the positive reactions are circled.
The sum of these indices in each group of three reactions forms the code number. This code is entered into the computer package. The percentage figure shown against the organism name is the percentage share of the probability for that organism as a part of the total probabilities for all choices.
Miscellaneous Gram-negative bacilli – Weakly positive reactions are recorded as negative results.
The results of tests for oxidase, nitrate reduction and motility are included as part of the reaction pattern. Using the results obtained, from each group of three reactions a 9 nine digit code number is produced.
Quality control The overall performance of the system should be monitored by testing appropriate control strains. The following organisms are recommended for independent laboratory assessment. The following chart gives the expected results on the Microbact System after an hour incubation: The expected results is positive. Some bacterial strains may have atypical biochemical reactions due to unusual nutritional requirements or mutations and may be difficult to identify.
Reactions obtained using the Microbact System may differ from published results using other substrate formulations. Prolonged incubation, insufficient incubation, improper filling of wells, or inadequate inoculum may lead to false results.
Species with low frequency of occurrence require additional testing. The interpretation of mathematically calculated identification results requires trained clinical personnel who should use judgement and knowledge in conjunction with the following information before accepting the ID of an organism: Gram-stain, colonial morphology, source of isolate, percent probability degree of separationtests against, additional test indications and results, frequency of ID choice and antibioGram.
A Gram-stain and oxidase test should be performed prior to set-up of tests. In addition, motility and nitrate test should be performed for miscellaneous Gram-negative bacilli. Twelve substrates provide insufficient data to speciate within this group as a single aberrant reaction may result in an incorrect identification.
The lysine and ornithine decarboxylase reactions should be carefully interpreted. Motility and DNase tests are recommended for further speciation of this group. The inclusion of a 12B strip is strongly advised.
If further speciation is required for Yersinia spp. Identification of Bacteria by Computer: General Aspects and Perspectives. Thomas, Veterinary Microbiology. Biochemical Characterisics of Enterotoxigenic Aeromonas sp. Material Safety Data Sheet. Microbact Biochemical Identification Kits. Materials Required but not provided The following materials may be required but are not provided: When Actinobacillus or Pasteurella spp. Well 1 lysine Well 2 galerke Well 3 H 2 S.
Green or blue is positive reaction. Bromothymol blue indicates formation of the specific amine cadaverine. Green should be regarded as a gzlerie reaction.