Setting up an API20E Biochemical Test StripPick up a single isolated colony (from a pure culture) and make a suspension of it in sterile distilled water.Take the API20E biochemical test strip which contains dehydrated bacterial media/biochemical reagents in 20 separate compartments. API20E biochemical test strip is commercially available. (Bacteria will react with them and will give different colors which will help to identify bacteria to the species level).Take a Pasteur pipette and fill up (up to the brim) these compartments with the bacterial suspension.Add sterile oil into the ADH, LDC, ODC, H2S, and URE compartments.Put some drops of water in the tray and put the API Test strip and close the tray.Mark the tray with an identification number (patient ID or organism ID), date, and your initials.Incubate the tray at 37oC for 18 to 24 hours.Results and InterpretationFor 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.Add the following reagents to these specific compartmentsTDA: Put one drop of ferric chlorideIND: Put one drop of Kovacs reagentVP: 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).Get the API reading scale (color chart)Mark each test as positive or negative on the lid of the trayThe wells are marked off into triplets by black triangles, for which scores are allocated as follows: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.Numbering in API 20E Test StripThe profile for this combination of reactions is therefore 7031645 (7 digit code)Identify the organism by using API catalog or apiwebVIDEO: Reading an API20E using the online database Identify the organism using APIWEB:Start Google Chrome or Firefox web browserGo to: Login: your login namePassword: your passwordSelect the correct test (e.g. API 20E).Enter the numerical profile to obtain the identity.Record the identity along with comments (% ID and T value) on the results sheet.Tweetwindow.twttr=(function(d,s,id)(document,"script","twitter-wjs"));lang:en_USEmail "@context":"http:\/\/schema.org\/","@type":"BlogPosting","name":"API 20E Test System: Results and Interpretations","url":"https:\/\/microbeonline.com\/api-20e-test-system-introduction-procedure-results-interpretations\/","articleBody":"API (Analytical Profile Index) 20E is a biochemical panel from bioMerieux,\u00a0Inc. for identifying and differentiating members of the family Enterobacteriaceae. The plastic strip holds twenty mini-test chambers containing dehydrated media having chemically defined compositions for each test. \n\n\n\nA bacterial suspension is used to rehydrate each of the wells. After incubation, some of the wells will have color changes due to pH differences: others produce end products that must be identified with reagents. A profile number is determined from the sequence of positive and negative test results, then looked up in a codebook to identify the bacterial isolate. \n\n\n\nAPI 20 E Biochemical Test Strip\n\n\n\nThese include:\n\n\n\nONPG: test for \u03b2-galactosidase enzyme by hydrolysis of the substrate o-nitrophenyl-b-D-galactopyranosideADH: decarboxylation of the amino acid arginine by arginine dihydrolaseLDC: decarboxylations of the amino acid lysine by lysine decarboxylaseODC: decarboxylations of the amino acid ornithine by ornithine decarboxylaseCIT: utilization of citrate as only carbon sourceH2S: production of hydrogen sulfideURE: test for the enzyme ureaseTDA (Tryptophan deaminase): detection of the enzyme tryptophan deaminase: Reagent to put- Ferric Chloride.IND: Indole Test-production of indole from tryptophan by the enzyme tryptophanase. Reagent- Indole is detected by the addition of Kovac's reagent.VP: the Voges-Proskauer test for the detection of acetoin (acetyl methylcarbinol) produced by fermentation of glucose by bacteria utilizing the butylene glycol pathwayGEL: test for the production of the enzyme gelatinase which liquefies gelatinGLU: fermentation of glucose (hexose sugar)MAN: fermentation of mannose (hexose sugar)INO: fermentation of inositol (cyclic polyalcohol)SOR: fermentation of sorbitol (alcohol sugar)RHA: fermentation of rhamnose (methyl pentose sugar)SAC: fermentation of sucrose (disaccharide)MEL: fermentation of melibiose (disaccharide)AMY: fermentation of amygdalin (glycoside)ARA: fermentation of arabinose (pentose sugar)\n\n\n\nSetting up an API20E Biochemical Test Strip\n\n\n\nPick up a single isolated colony (from a pure culture) and make a suspension of it in sterile distilled water.Take the API20E biochemical test strip which contains dehydrated bacterial media\/biochemical reagents in 20 separate compartments. API20E biochemical test strip is commercially available. (Bacteria will react with them and will give different colors which will help to identify bacteria to the species level).Take a Pasteur pipette and fill up (up to the brim) these compartments with the bacterial suspension.Add sterile oil into the ADH, LDC, ODC, H2S, and URE compartments.Put some drops of water in the tray and put the API Test strip and close the tray.Mark the tray with an identification number (patient ID or organism ID), date, and your initials.Incubate the tray at 37oC for 18 to 24 hours.\n\n\n\nResults and Interpretation \n\n\n\nFor 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.Add the following reagents to these specific compartments TDA: Put one drop of ferric chlorideIND: Put one drop of Kovacs reagentVP: Put one drop of 40 % KOH (VP reagent 1) & One drop of VP Reagent 2 (\u03b1-Naphthol) (you have to wait for 10 minutes before telling it negative). Get the API reading scale (color chart) Mark each test as positive or negative on the lid of the trayThe wells are marked off into triplets by black triangles, for which scores are allocated as follows: 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. \n\n\n\nNumbering in API 20E Test Strip\n\n\n\nThe profile for this combination of reactions is therefore 7031645 (7 digit code)Identify the organism by using API catalog or apiwebVIDEO: Reading an API20E using the online database\u00a0Identify the organism using APIWEB: Start Google Chrome or Firefox web browserGo to: https:\/\/apiweb.biomerieux.com Login: \u00a0your login\u00a0namePassword: your passwordSelect the correct test (e.g. API 20E).Enter the numerical profile to obtain the identity.Record the identity along with comments (% ID and T value) on the results sheet.","headline":"API 20E Test System: Results and Interpretations","author":"@type":"Person","name":"Acharya Tankeshwar","url":"","datePublished":"2015-05-06","mainEntityOfPage":"True","dateModified":"October 6, 2022","image":"@type":"ImageObject","url":"https:\/\/microbeonline.com\/wp-content\/uploads\/2015\/05\/API-20-E-1024x140.jpg","height":88,"width":640,"publisher":"@context":"http:\/\/schema.org\/","@type":"Organization","name":"Microbe Online","logo":"@type":"ImageObject","url":"https:\/\/microbeonline.com\/wp-content\/uploads\/2022\/08\/microbeonline-logo-300x135.png","height":600,"width":60Acharya TankeshwarHello, thank you for visiting my blog.I am Tankeshwar Acharya. Blogging is my passion. As an asst. professor, I am teaching microbiology and immunology to medical and nursing students at PAHS, Nepal. I have been working as a microbiologist at Patan hospital for more than 10 years.if(typeof ez_ad_units!='undefined')ez_ad_units.push([[300,250],'microbeonline_com-large-mobile-banner-1','ezslot_7',879,'0','0']);__ez_fad_position('div-gpt-ad-microbeonline_com-large-mobile-banner-1-0');Load CommentsRecent Postslink to API and RAPID ID For Microbial IdentificationAPI and RAPID ID For Microbial IdentificationBioMérieux's API identification products are test kits for the identification of Gram-positive bacteria, Gram-negative bacteria, and yeast. The kits include strips that contain up to 20 miniature...Continue Readinglink to Biochemical Tests for Bacterial IdentificationBiochemical Tests for Bacterial IdentificationIn this blog post, I am sharing information about the standard, conventional biochemical tests and their primary uses in the Microbiology laboratory for identifying pathogens of interest. Some of the...Continue Readingif(typeof ez_ad_units!='undefined')ez_ad_units.push([[300,250],'microbeonline_com-box-1','ezslot_4',200,'0','0']);__ez_fad_position('div-gpt-ad-microbeonline_com-box-1-0');report this adAbout UsMicrobeonline.com is an online guidebook on Microbiology, precisely speaking, Medical Microbiology. This blog shares information and resources about pathogenic bacteria, viruses, fungi, and parasites. We are trying our best to make this site user-friendly and resourceful with timely/updated information about each pathogen, disease caused by them, pathogenesis, and laboratory diagnosis.
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