AN MicroPlate - Oxoid

Various methods have different numbers and types of
organisms within their database. Figure 2 presents several
popular kit-based methods. The Biolog AN MicroPlate has a
much larger number of tests, which provides greater
fingerprint discrimination and a larger database.
Manufacturer Number of
Anaerobic Species
in Database
Biolog, Inc
Microlog TM
bioMérieux
Vitex ®
bioMérieux
API 20A ®
BBL ® Crystal TM
Innovative
Diagnostic
Systems, Inc.
RapID ANA II
Number of Tests
Used for Detection
359 95
84 28
77 20
107 28
96 18
FIGURE 2. Comparison of Commercial
Test Kits for Anaerobes
Beyond the number of tests provided to detect an unknown,
some methods rely primarily on fermentation of sugars. This
approach to identification does not provide the necessary
environment for every organism of interest. Many bacteria
cannot utilize sugars via a fermentative process and are missed
or react weakly with these methods.
The state of anaerobic taxonomy further exacerbates the
limitations of other methods. The taxonomy of anaerobic
bacteria is based on much less information and is not as
developed as aerobic taxonomy. For example, the
classification of Clostridium botulinum is based on the
organism’s ability to produce one or more forms of botulinum
neurotoxin. Five morphologically distinct organisms have
been shown to produce this neurotoxin. According to current
anaerobic taxonomy all five of these organisms should be
classified (identified) as C. botulinum. Strains of these species
which do not produce the toxin, are not classified as nontoxigenic
C. botulinum, but would be called, for example C.
sporogenes. If this same criterion were applied to aerobic
taxonomy then all Shiga toxin producing species of enteric
bacteria would be assigned to the genus Shigella.
Anaerobe Identification Test Panel
PROCEDURE FOR USING AN MICROPLATES
The procedure is fast and simple, involving only 5 steps, and
requiring only 2 to 3 minutes hands-on time per sample.
1) A pure culture of a bacterium is grown on a Biolog
Universal Anaerobe agar plate (Biolog Catalog #
70007 for a 500g jar or 71007 for pre-poured plates)
in either an anaerobic chamber or anaerobic jar until
enough growth is present to prepare a suspension.
2) The bacteria are swabbed from the surface of the agar
plate, and suspended to a specified density in AN
Inoculating Fluid (Biolog Catalog # 72007). This
step may be performed in an anaerobic chamber.
3) 100 µl of bacterial suspension is pipetted into each
well of the AN MicroPlate (Biolog Catalog # 1007)
under aerobic conditions.
4) The MicroPlate is incubated in an anaerobic jar with
a hydrogen free anaerobic atmosphere at 35 o C for
20-24 hours. (A hydrogen free anaerobic atmosphere
can be produced by either the Oxoid AnaeroGen or
the Mitsubishi AnaeroPak System, Pack-Anaero.)
5) The MicroPlates are removed from the anaerobic
container and read either visually or with the Biolog
MicroStation Reader.
ANAEROBE DATABASE
The Biolog AN database has the largest database of any kitbased
method. This superior product will be an invaluable
addition to your anaerobic laboratory. Some highlights are:
o Bifidobacterium, usually called Bifidus, which could
not formerly be speciated by ID kits, can now be
identified with this product.
o The genera Pectinatus and Megasphaera are not
covered in other anaerobic test kits but can be
identified with the MicroLog AN database.
o The MicroLog database has 43 Lactobacillus spp.,
and over 70 species of lactic acid bacteria. Other kit
based methods cover approximately 7 species.
For more information, contact us using the information below.
3938 Trust Way Hayward, CA 94545 Telephone: 510-785-2564 Fax: 510-782-4639 www.biolog.com
05/01

AN MicroPlate
Anaerobe Identification Test Panel
A1 A2 A3 A4 A5 A6 A7 A8 A9 A10 A11 A12
N-Acetyl- N-Acetyl- N-Acetyl- Adonitol Amygdalin D-Arabitol Arbutin D-Cellobiose α-Cyclodextrin β-Cyclodextrin Dextrin
Water D-Galactosamine D-Glucosamine D-Mannosamine
B1 B2 B3 B4 B5 B6 B7 B8 B9 B10 B11 B12
Dulcitol I-Erythritol D-Fructose L-Fucose D-Galactose D-Galacturonic Gentiobiose D-Gluconic D-Glucosaminic α-D-Glucose Glucose-1- Glucose-6-
Acid Acid Acid Phosphate Phosphate
C1 C2 C3 C4 C5 C6 C7 C8 C9 C10 C11 C12
Glycerol D,L-α-Glycerol m-Inositol α-D-Lactose Lactulose Maltose Maltotriose D-Mannitol D-Mannose D-Melezitose D-Melibiose 3-Methyl-
Phosphate D-Glucose
D1 D2 D3 D4 D5 D6 D7 D8 D9 D10 D11 D12
α-Methyl- β-Methyl- α-Methyl- β-Methyl- Palatinose D-Raffinose L-Rhamnose Salicin D-Sorbitol Stachyose Sucrose D-Trehalose
D-Galactoside D-Galactoside D-Glucoside D-Glucoside
E1 E2 E3 E4 E5 E6 E7 E8 E9 E10 E11 E12
Turanose Acetic Acid Formic acid Fumaric Acid Glyoxylic Acid α- Hydroxy- β- Hydroxy- Itaconic Acid α-Ketobutyric α-Ketovaleric D,L-Lactic L-Lactic Acid
butyric Acid butyric Acid Acid Acid Acid
F1 F2 F3 F4 F5 F6 F7 F8 F9 F10 F11 F12
D-Lactic Acid D-Malic Acid L-Malic Acid Propionic Acid Pyruvic Acid Pyruvic Acid D-Saccharic Succinamic Succinic Acid Succinic Acid M-Tartaric Acid Urocanic Acid
Methyl Ester Methyl Ester Acid Acid Mono-Methyl
Ester
G1 G2 G3 G4 G5 G6 G7 G8 G9 G10 G11 G12
Alaninamide L-Alanine L-Alanyl- L-Alanyl- L-Alanyl- L-Asparagine L-Glutamic Acid L-Glutamine Glycyl- Glycyl- Glycyl- Glycyl-
L-Glutamine L-Histidine L-Threonine L-Aspartic Acid L-Glutamine L-Methionine L-Proline
H1 H2 H3 H4 H5 H6 H7 H8 H9 H10 H11 H12
L-Methionine L-Phenylalanine L-Serine L-Threonine L-Valine L-Valine plus 2'-Deoxy Inosine Thymidine Uridine Thymidine-5'- Uridine-5'-
L-Aspartic Acid Adenosine Mono-phosphate Mono-phosphate
INTRODUCTION
The Biolog AN MicroPlate (Figure 1) is designed for
identification of a very wide range of anaerobic bacteria,
including the genera Bifidobacterium, Clostridium,
Eubacterium, Fusobacterium, Lactobacillus, Lactococcus,
Megasphaera, Pectinatus, Pediococcus, Peptostreptococcus,
Propionibacterium and Weissella. These genera are important
in industrial and environmental applications, especially the
food industry where they are responsible for both food
production and food spoilage. Some clinical species are not
currently in our database. These will be added when they
meet our performance criteria. The AN MicroPlate employs
the same redox chemistry used in the Biolog GP2 and GN2
MicroPlates. This chemistry, based on reduction of
tetrazolium, responds to the process of metabolism (oxidation
of substrates) rather than to metabolic by-products (e.g. acid).
Biolog’s universal chemistry works with any carbon source
and greatly simplifies the testing process, as no color
developing chemicals need to be added after incubation.
AN MICROPLATE
The Anaerobe Database contains over 350 taxa of anaerobic
bacteria including over 70 species of lactic acid bacteria.
FIGURE 1. Carbon Source in AN MicroPlate
The Biolog AN MicroPlate performs 95 discrete tests
simultaneously and gives a characteristic reaction pattern
called a “metabolic fingerprint”. These fingerprint reaction
patterns provide a vast amount of information conveniently
contained on a single Biolog MicroPlate. The patterns are
compared using Biolog MicroLog database software to give
an identification.
Other anaerobic kit-based identification methods rely on fewer
tests to perform identifications. Therefore an anaerobic
organism that was previously characterized by another method
may yield an identification result that differs from the Biolog
AN MicroPlate & Database. This difference may be due to
several factors. When determining the validity of an
anaerobic identification result from the Biolog AN MicroPlate
and another method, consider the following:
o The MicroLog System bases its identification on a
larger number of tests
o The MicroLog System covers more species
o The taxonomy of many anaerobic genera is poorly
defined