Gram Negative Identification Test Panel GN2 Microplate™

Gram Negative Identification Test Panel

A1 A2 A3 A4 A5 A6 A7 A8 A9 A10 A11 A12 Water α-Cyclodextrin Dextrin Glycogen Tween 40 Tween 80 N-Acetyl-D- N-Acetyl-D- Adonitol L-Arabinose D-Arabitol D-Cellobiose galactosamine glucosamine

B1 B2 B3 B4 B5 B6 B7 B8 B9 B10 B11 B12 α α i-Erythritol D-Fructose L-Fucose D-Galactose Gentiobiose α-D-Glucose m-Inositol α-D-Lactose Lactulose Maltose D-Mannitol D-Mannose

C1 C2 C3 C4 C5 C6 C7 C8 C9 C10 C11 C12 D-Melibiose β-Methyl- D-Psicose D-Raffinose L-Rhamnose D-Sorbitol Sucrose D-Trehalose Turanose Xylitol Methyl Mono-Methyl- D-Glucoside Pyruvate Succinate

D1 D2 D3 D4 D5 D6 D7 D8 D9 D10 D11 D12 Acetic Acid Cis-Aconitic Citric Acid Formic Acid D-Galactonic D-Galacturonic D-Gluconic D-Glucosaminic D-Glucuronic α-Hydroxy β-Hydroxy γ-Hydroxy Acid Acid Lactone Acid Acid Acid Acid Butyric Acid Butyric Acid Butyric Acid

E1 E2 E3 E4 E5 E6 E7 E8 E9 E10 E11 E12 p-Hydroxy Itaconic Acid α-Keto Butyric α-Keto Glutaric α-Keto Valeric D,L-Lactic Acid Malonic Acid Propionic Acid Quinic Acid D-Saccharic Sebacic Acid Succinic Acid Phenylacetic Acid Acid Acid Acid Acid

F1 F2 F3 F4 F5 F6 F7 F8 F9 F10 F11 F12 Bromo Succinamic Glucuronamide L-Alaninamide D-Alanine L-Alanine L-Alanyl- L-Asparagine L-Aspartic Acid L-Glutamic Glycyl-L- Glycyl-L- Succinic Acid Acid glycine Acid Aspartic Glutamic Acid Acid G1 G2 G3 G4 G5 G6 G7 G8 G9 G10 G11 G12 L-Histidine Hydroxy-L- L-Leucine L-Ornithine L- L-Proline L-Pyroglutamic D-Serine L-Serine L-Threonine D,L-Carnitine γ-Amino Butyric Proline Phenylalanine Acid Acid

H1 H2 H3 H4 H5 H6 H7 H8 H9 H10 H11 H12 Urocanic Acid Inosine Uridine Thymidine Phenyethylami Putrescine 2-Aminoethanol 2,3-Butanediol Glycerol D,L-α-Glycerol Glucose-1- Glucose-6- ne Phosphate Phosphate Phosphate

FIGURE 1. Carbon Sources in GN2 MicroPlate

INTRODUCTION GN2 MICROPLATE

The Biolog GN2 MicroPlate (Figure 1) is designed for The Biolog GN2 MicroPlate performs 95 discrete tests identification and characterization of a very wide range of simultaneously and gives a characteristic reaction pattern aerobic gram-negative bacteria. Biolog’s MicroPlates and called a “metabolic fingerprint”. These fingerprint reaction databases were first introduced in 1989, employing a novel, patterns provide a vast amount of information conveniently patented redox chemistry. This chemistry, based on reduction contained in a single Biolog MicroPlate. The metabolic of tetrazolium, responds to the process of metabolism (i.e. fingerprint patterns are compared and identified using the respiration) rather than to metabolic by-products (e.g. acid). MicroLog™ database software. Biolog’s chemistry works as a universal reporter of Other aerobic kit-based identification methods rely on a much metabolism and simplifies the testing process as color smaller number of tests. Consequently, the significant developing chemicals do not need to be added. Since the GN2 limitation of these products is the limited number of species MicroPlate is not dependent upon growth to produce and organism types that they can identify. Furthermore, these identifications, it provides superior capability for all types of products were designed to address the needs of routine gram negative organisms: fermenters, non-fermenters, and clinical/hospital testing. The Biolog GN2 MicroPlate was fastidious organisms all are identified with a single panel. The designed to address the needs of a much wider range of users database for the GN2 MicroPlate is now over 500 species. It including environmental testing labs and animal and plant is by far the largest kit-based identification database available. disease labs as well as clinical reference labs.

Gram Negative Identification Test Panel

There are approximately 4,000 named bacterial species and The GN2 MicroPlate has demonstrated accuracy comparable this is just a fraction of the total number of species in the to molecular methods and without the expense. Figure 3, environment. The MicroLog System provides the unique provides representative results from an independent study with feature of user defined custom databases. If an organism is non-routine isolates compared to a molecular method: outside the MicroLog database, the user can save the pattern to a custom database for future reference. If the organism is % Correct to the Biolog, Inc Molecular Method isolated again, the laboratory will have the pattern saved Species Level MicroLog instead of simply getting a “no ID”. Some other methods provide supplemental off-line tests for use alongside the Fermenters 80% 72% identification panel. This approach is inconvenient and does Nonfermenters 88% 100% not produce an expanded pattern library. Overall 85% 89%

An identification from the Biolog GN2 MicroPlate is superior Figure 3: Comparison of Phenotypic and Genotypic 1 to less precise methods, because: Techniques

o The MicroLog System bases its identification on a PROCEDURE FOR USING GN2 MICROPLATES larger number of tests. There are over 4 x 1028

possible patterns from a single MicroPlate The test procedure is fast and simple, involving only 5 steps, o The MicroLog System covers far more species and requiring only 2 to 3 minutes hands-on time per sample. 1) A pure culture of a bacterium is grown on a Biolog o Older methods were developed to detect routine Universal Growth w/5% Sheep Blood agar plate clinical pathogens, and do not adequately identify (Biolog catalog #70101 for a 500g jar of dehydrated other important organisms such as: Acinetobacter powder.) spp., Aeromonas spp., Bordetella spp., Haemophilus 2) The bacteria are swabbed from the surface of the agar spp., Pseudomonas spp., Vibrio spp., and Yersinia plate, and suspended to a specified density in GN/GP spp. Inoculating Fluid (Biolog catalog # 72101). Various methods have different numbers and types of 3) 150 µl of bacterial suspension is pipetted into each organisms within their database. Figure 2, compares several well of the GN2 MicroPlate (Biolog catalog # 1011). popular kit-based methods. The Biolog GN2 MicroPlate has a 4) The MicroPlate is incubated at 30° or 35o C much larger number of tests, which provides greater (depending upon the nature of the organism) for 4-24 fingerprint discrimination and a larger database. hours. 5) The MicroPlates are read either visually or with the Manufacturer Number of Aerobic Number of Tests Biolog MicroStation or OmniLog System, Species in Database Used for Detection compared to the GN Database (Biolog catalog # 22401A), and a result is determined. Biolog, Inc 501 95 MicroLog REFERENCES: BioMérieux 104 29 Vitek® GNI+ [1] Comparison of Phenotypic and Genotypic Techniques of Identification of Unusual Aerobic bioMérieux API ~180 20 ® Pathogenic Gram Negative Bacilli. Y.W. Tang, N.M. 20E & NFT Ellis, M.K. Hopkins, D.E. Dodge, and D.H. Persing, BBL® Crystal™ ~105 28 Journal of Clinical Microbiology, December 1998, p. 3674-3679 FIGURE 2. Comparison of Commercial Test Kits for Gram Negative Organisms CONTACT INFORMATION In addition to a limited number of tests used to identify an unknown, some methods rely primarily on fermentation of The Biolog Microbial Identification/Characterization System sugars. This approach does not provide the necessary will be an invaluable addition to your microbiology environment for every organism of interest. Many bacteria laboratory. cannot utilize sugars via a fermentative process and react weakly or not at all with these methods. The larger number For more details, contact us using the information below: and more diverse range of tests in the GN2 MicroPlate provide for greater accuracy and precision.

3938 Trust Way Hayward, CA 94545 Telephone: 510-785-2564 Fax: 510-782-4639 www.biolog.com

Part# OOA 003, Rev. A, Mar. 2001