The world leader in serving science Proprietary & Confidential
Julie Elston Global Product Manager, Immunodiagnostics and Product Training Manager
Selling the Portfolio
2 Proprietary & Confidential
• Industrial Workflow • Food testing
• Media, DryBags, Precis • Confirmation – biochemical ID and latex agglutination
• Water Testing
• Media and confirmation
• Pharmaceutical • Environmental monitoring – TWIP and *NEW* Contact Plate Applicator
Agenda
3 Proprietary & Confidential
• Extensive product range
• Our core business is manual microbiology
• We are moving forward with technological advances
• BUT...... ......we must protect our market share and we must protect our customer base
in preparation for future developments
• There are still many opportunities , especially cross-selling products from the customer’s workflow
Our portfolio
The world leader in serving science Proprietary & Confidential
Industrial Workflow
Sample Collection
Fisher Scientific
Sample Enrichment
/Dilution
Sample Preparation/Purification
Detection Confirmation/ Identification Further ID
5 Proprietary & Confidential
• Food Testing
• Water Testing
• Pharmaceutical testing
Industrial workflow
6 Proprietary & Confidential
Sample Collection
Fisher Scientific
Sample Enrichment
/Dilution
Sample Preparation/Purification
Detection Confirmation/ Identification Further ID
Food Testing – laboratory workflow
7 Proprietary & Confidential
Food Testing – What do people look for and why?
• Pathogens • To ensure that food is safe and meets legislation
• Toxins • Some organisms such as Salmonella produce dangerous toxins
• Indicator organisms • Counts, TVC, E. coli, coliforms, Enterobacteriaceae A change in expected number indicates that something in the process has
gone wrong or changed
• The presence of specific organisms may indicate the increased likelihood of certain pathogens being present (monitoring normal levels not just presence absence)
Listeria spp. monitoring could indicate L. monocytogenes levels and E. coli monitoring could indicate E. coli O157
• Spoilage organisms • Organisms that will cause food to go off or cause undesirable traits e.g. Yeast & Moulds
8 Proprietary & Confidential
Microbiological Food Testing
1. Pre-enrichment
2. Selective Enrichment
3. Selective Plating
4.Confirmation Tests
9 Proprietary & Confidential
Pre-enrichment
• Helps growth of target organisms that may be stressed e.g. drying, low pH and high temperature
• Helps grow sufficient numbers of bacteria to be detected on media
• No select agents (antibiotics) as this may harm the target organism
• Competitor organisms grow as well
1. Pre-enrichment
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Selective enrichment
• Contains selective agents to kill non-target organisms • Increases growth of the target organism
2. Selective Enrichment
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Selective Plating
• More selective agents included to inhibit the growth of non-target organisms that grew in selective enrichment
• Allows differentiation of target organism usually by • Colour of colonies • Clear zones
3. Selective
Plating
12 Proprietary & Confidential
Confirmation of positives • Confirms that you have isolated your target
organism
• A wide selection of tests can be performed, according to target organism
• Physical tests • Gram stain, motility, shape
• Biochemical tests • Sugar metabolism • Oxidase • Catalase
• Immunological tests
• Agglutination of homologous antibody/antigen
4.Confirmation Tests
The world leader in serving science Proprietary & Confidential
Salmonella Précis and Listeria Précis Workflows
14 Proprietary & Confidential
2 days
Salmonella Precis
ENRICHMENT ONE Broth-Salmonella
PLATING Brilliance™ Salmonella
CONFIRMATION Oxoid Salmonella Latex
or ISO 6579 methods
15 Proprietary & Confidential
Listeria Precis Workflow
2 days
ENRICH ONE Broth-Listeria
PLATE Brilliance™ Listeria
CONFIRM O.B.I.S. mono
or ISO 11290
The world leader in serving science Proprietary & Confidential
Innovative format and workflow solution for efficient food pathogen testing
DryBags
17 Proprietary & Confidential
Automated Salmonella Précis Preparation
Peristaltic Pump
ONE Broth Dry-Bag
Automated Diluter
Brilliance Salmonella
Salmonella Latex
15 – 30 mins 1 sample/min 24 hours/48
hours 15 mins
Sample Collection
Sample Enrichment
Sample Purification Detection Confirmation ID/Charact
erization
2 days
18 Proprietary & Confidential
Automated Précis Preparation
Peristaltic Pump
ONE Broth Dry-Bag
Automated Diluter
Brilliance Listeria
O.B.I.S Listeria
15 – 30 mins 1 sample/min 24 hours/48
hours 15 mins
Sample Collection
Sample Enrichment
Sample Purification Detection Confirmation ID/Charact
erization
2 days
The world leader in serving science Proprietary & Confidential
Food Microbiology Confirmation Methods
20 Proprietary & Confidential
Customers
Food Laboratories… • World population is growing and consuming more, leading to an increase in the number of food microbiology tests carried out each year
• Higher demands and expectations as well as increased levels of regulation as governments attempt to ensure that manufacturers meet minimum safety levels for food
• QC laboratory manager is the key person in deciding which tests to run, where they are done, what methods are used and the frequency of testing
Sample Collection
Sample Enrichment
/Dilution
Sample Prep/
Purification Detection Confirmation Identification/
Characterization
21 Proprietary & Confidential
Biochemical Identification - What it is and how it works
Sample Collection
Sample Enrichment
/Dilution
Sample Prep/
Purification Detection Confirmation Identification/
Characterization
22 Proprietary & Confidential
Thermo Scientific™ Remel™ Spot Tests
Single-substrate reagents for rapid identification of microorganisms • Skilled microbiologists can identify the majority of isolates seen routinely using
colony and Gram-stain morphology, and one-step biochemical spot tests • E. coli can be identified based on growth and lactose fermentation of MacConkey
Agar, indole positivity, oxidase negativity, and beta glucuronidase positivity (MUG) within one hour at low cost
Sample Collection
Sample Enrichment
/Dilution
Sample Prep/
Purification Detection Confirmation Identification/
Characterization
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Rapid Spot Test range
Product # Product Name Organism ID
R21132 Microdase Disk Modified oxidase test for differentiation of staphylococci from micrococci
R21135 MUG Disk For presumptive identification of Escherichia coli by fluorogenic assay
R211667 Nitrocefin Disk Haemophilus spp., N. gonorrhoeae, M. catarrhalis, staphylococci, enterococci, and anaerobes
R21134 Novobiocin Disk For differentiation of Staphylococcus saprophyticus from other coagulase-negative staphylococci
R211357 PRO Disk w/Reagent For identification of yeast and anaerobic microorganisms primarily Candida albicans and Clostridium difficile
R211172 PYR Disk w/Reagent For presumptive identification of enterococci, Lancefield group A streptococci, and E. coli
R21138 PYR/Esculin Disk For presumptive identification of group A streptococci and enterococci
R21137 Pyridoxal Disk For demonstration of growth by nutritionally deficient streptococci
R21152 Urea-PDA Disk For demonstration of urease activity and phenylalanine deamination in Enterobacteriaceae
R21115 ALA Disk For demonstration of porphyrin synthesis in Haemophilus spp.
R261605 Beta Lactam Disk Detection of beta-lactamase production in Haemophilus spp., N. gonorrhoeae, and staphylococci
R21120 Beta Lysin Disk For detection of CAMP factor by group B streptococci
R21122 Bile Disk Screening test to separate Bacteroides fragilis from other gram-negative anaerobic bacilli
R21128 Caffeic Acid Disk For demonstration of phenol oxidase production by Cryptococcus neoformans
R21121 Catarrhalis Test Disk For rapid detection of butyrate esterase for presumptive identification of Moraxella catarrhalis
R211820 Gram-Sure Rapid test disk for differentiation of aerobic gram-positive and gram-negative rods or coccobacilli
R21085 Hippurate Disk Detect hydrolysis of sodium hippurate by group B streptococci, G. vaginalis, and C. jejuni
R21087 Indoxyl Acetate Disk For rapid identification and differentiation of Campylobacter, Wolinella, and Helicobacter
R21129 LAP Disk For the rapid detection of leucine aminopeptidase activity in streptococci and related genera
R30168501 LAP Disk w/Reagent Differentiation Aerococcus/Leuconostoc from Streptococcus/Enterococcus/Lactococcus/Pediococcus
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Oxoid Biochemical Identification Systems
• OBIS PYR (ID0580M) - determination of PYRase activity in streptococci and Citrobacter • OBIS Mono (ID0600M) - differentiation of L. monocytogenes from other Listeria species • OBIS Campy (ID0800M) - differentiation of Campylobacteriaceae from other Gram negative
organisms • OBIS Salmonella ( ID0570M) - to differentiate Salmonella spp. from similar organisms • OBIS albicans (ID0700M) - differentiation of Candida albicans from other yeast species
• Position as quick, safe and simple, confirmatory tests • Results in 2 – 12 minutes from testing colony • Reduce the number of more expensive high level identifications • Quickly screen out negatives
Thermo Scientific™ O.B.I.S.™
Sample Collection
Sample Enrichment
/Dilution
Sample Prep/
Purification Detection Confirmation Identification/
Characterization
25 Proprietary & Confidential
O.B.I.S. Kit General Protocol
1. Carry out orientation tests such as catalase, oxidase and/or Gram stain 2. Smear organism into reaction area(s) of card 3. Moisten reaction area of card 4. Allow to react (time and temperature depend on test kit; 30 seconds – 1
hour) 5. Add developing solution to reaction area(s) 6. Wait for reaction to take place (20 seconds) 7. Look for colour reactions and record results 8. Determine if further identification is required
Refer to pack insert for exact methodology of each kit
Sample Collection
Sample Enrichment
/Dilution
Sample Prep/
Purification Detection Confirmation Identification/
Characterization
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• AOAC Official Method – first choice of system in food
• 15 biochemical tests detecting specific enzymes and/or metabolic end products
• Self-contained test system with reagent impregnated discs
• R38370 – Micro-ID Listeria ID System • 4 hour screen for L. monocytogenes
• Positive esculin and rhamnose disks
• 24 hour incubation for species identification
• R38145 – Micro-ID Identification System • 4 hour incubation • Enterobacteriaceae identifications
Thermo Scientific™ Remel™ MICRO-ID™ Identification System
Sample Collection
Sample Enrichment
/Dilution
Sample Prep/
Purification Detection Confirmation Identification/
Characterization
27 Proprietary & Confidential
Thermo Scientific™ Remel™ MICRO-ID™ LISTERIA
• Test Gram-postive, oxidase-negative, catalase-positive colonies • Prepare organism suspension to McFarland No. 1 • Inoculate each well • Close lid and incubate at 35 – 37oC for 24 hours • After 4 hours, l. monocytogenes suspsected if esculin (E) and rhamnose (RHAM) are
positive • After 24 hours add reagent to VP disk • Rotate device to moisten disks. Read and record colour change
Sample Collection
Sample Enrichment
/Dilution
Sample Prep/
Purification Detection Confirmation Identification/
Characterization
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• Self-contained biochemical test system
• MB1128A – Microbact Listeria 12L • 11 carbohydrate utilization test and 1 rapid microhaemolysis test
• Identification in as little as 4 hours
• No prior subculturing on non-selective medium, can be used with chromogenic or selective agars
• MB1074A – Microbact GNB Kit (several configurations)
• 24 tests total available in strips of 12 tests which can be used independently or together
• MB1561A – Microbact Staphylococcal 12S • 12 tests based on sugar utilization and enzyme detection substrates
Thermo Scientific™ Oxoid™ Microbact™
Sample Collection
Sample Enrichment
/Dilution
Sample Prep/
Purification Detection Confirmation Identification/
Characterization
29 Proprietary & Confidential
• Test Gram-postive, oxidase-negative, catalase-positive colonies
• Prepare suspending medium • Suspend single isolated colony for 24 hr test • Prepare McFarland 0.5 Standard for 4 hour test
• Inoculate each well with 4 drops of suspension • Add 1 drop of haemolysin reagent to Well 12 • Replace lid and incubate at 35 – 37oC for 4 or 24 hours • Remove lid and read and record colour change
Thermo Scientific™ Oxoid™ Microbact™ Listeria 12L
Sample Collection
Sample Enrichment
/Dilution
Sample Prep/
Purification Detection Confirmation Identification/
Characterization
30 Proprietary & Confidential
Thermo Scientific™ Remel™ RapID™ Systems
• When a confirmation must be made, waiting for an ID panel to do its job can be frustrating
• Traditionally, manual, confirmatory ID panels provide results in 24 – 48 hours because they utilize carbohydrate fermentation reactions that are dependent on organism growth
RapID works differently
• It uses enzyme technology to reduce the time-to-results to 4 hours • Based on microbial degradation of specific substrates • Combination of conventional tests and single-substrate chromogenic tests • The advantage - provide a definitive answer to bacterial ID faster
Sample Collection
Sample Enrichment
/Dilution
Sample Prep/
Purification Detection Confirmation Identification/
Characterization
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RapID Principle
• Ideal for stand-alone testing or for complementing automated systems when testing for anaerobes, yeast, Neisseria-Haemophilus, Enterobacteriaceae, staphylococci, streptococci, Corynebacteria, non-fermenters, and urinary tract isolates
• Reaction cavities contain dehydrated reactants and the tray allows the simultaneous inoculation of each cavity with a predetermined amount of inoculum
• A suspension of the test organism in RapID Inoculation Fluid is used as the inoculum which rehydrates and initiates test reactions
• After incubation of the panel, each test cavity is examined for reactivity by noting the development of colour
• The resulting pattern of positive and negative test scores is used as the basis for identification of the test isolate by comparison of test results to reactivity patterns stored in a database
Sample Collection
Sample Enrichment
/Dilution
Sample Prep/
Purification Detection Confirmation Identification/
Characterization
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Features & benefits
Sample Collection
Sample Enrichment
/Dilution
Sample Prep/
Purification Detection Confirmation Identification/
Characterization
34 Proprietary & Confidential
• Simple biochemical tests help to address current lab issues • Best use of the laboratory’s resources • Assure quality test results • Reasonable cost
• Extensive range of single step biochemical tests
• Part of the end-to-end workflow solution
Spot Tests and O.B.I.S. Competitive Advantage
Sample Collection
Sample Enrichment
/Dilution
Sample Prep/
Purification Detection Confirmation Identification/
Characterization
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MICRO-ID™ Competitive Advantage
Sample Collection
Sample Enrichment
/Dilution
Sample Prep/
Purification Detection Confirmation Identification/
Characterization
• AOAC Official Method • First choice identification method for customers
• 4 hour Enterobacteriaceae identification
• 4 hour L. monocytogenes screen • 24 hour species level identification for all Listeria species
• Part of the end-to-end workflow solution
36 Proprietary & Confidential
Oxoid™ Microbact™ Competitive Advantage
Sample Collection
Sample Enrichment
/Dilution
Sample Prep/
Purification Detection Confirmation Identification/
Characterization
• 4 hour incubation for Listeria species
• 24 hours for Gram negative bacteria and Staphylococcus species
• Simple, strip format
• Can be an alternative to Micro-ID products
• Part of the end-to-end workflow solution
37 Proprietary & Confidential
RapID™ Systems Competitive Advantage
Sample Collection
Sample Enrichment
/Dilution
Sample Prep/
Purification Detection Confirmation Identification/
Characterization
38 Proprietary & Confidential
Latex Agglutination - What it is and how it works
Sample Collection
Sample Enrichment
/Dilution
Sample Prep/
Purification Detection Confirmation Identification/
Characterization
39 Proprietary & Confidential
Thermo Scientific™, Oxoid™ and Remel™ Latex Agglutination Kits
Sample Collection
Sample Enrichment
/Dilution
Sample Prep/
Purification Detection Confirmation Identification/
Characterization
• For the detection or identification of micro-organisms using immunological technology
• Polystyrene latex particles are sensitised with either a specific antigen • Ready-to-use reagents • Simple, easy to perform procedures • Rapid results • Particle agglutination
• Red blood cell agglutination • Latex agglutination
• wet latex • dry latex
• FDA 510K approved • CE marked
40 Proprietary & Confidential
Test Principles – Agglutination tests
The tests are always
• Rapid • Simple • Visual
Sample Collection
Sample Enrichment
/Dilution
Sample Prep/
Purification Detection Confirmation Identification/
Characterization
41 Proprietary & Confidential
Test Principles - Latex Agglutination
• For the detection or identification of micro-organisms or specific antibodies using immunological technology
• The polystyrene latex particles are sensitised with either a specific immunoglobulin or specific antigen
Key
Bacteria (antigen)
Latex particle
Y (sensitised with antibody)
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A positive latex agglutination reactions is indicated by the visual clumping of particles with clearing of the opaque background A negative latex agglutination reaction is indicated by a lack of clumping and the opaque background stays unchanged
Positive Negative
Agglutination Tests
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Food Agglutination Kits (wet)
Name No. of tests Code • Wellcolex E. coli O157:H7 50 R30959501/R30959601 • Wellcolex Colour Salmonella 50/200 R30858301/R30858302 • Staphaurex 120/400 R30859901/R30859902 • Staphaurex Plus 150/450 R30950102/R30950201 • Campylobacter Test Kit 50 DR0155M • Staphylase 100 DR0595A • E.coli O157 Kit 100 DR0620M • Staphytect Plus 100/500 DR0850M/B • PBP2I 50 DR0900A • Salmonella Test Kit 100 DR1108A • Listeria Test Kit 100 DR1126A
Sample Collection
Sample Enrichment
/Dilution
Sample Prep/
Purification Detection Confirmation Identification/
Characterization
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Food DrySpot Agglutination Kits
Name No. of tests Code • Staphytect Plus 120 DR0100M • E. coli O157 120 DR0120M
Sample Collection
Sample Enrichment
/Dilution
Sample Prep/
Purification Detection Confirmation Identification/
Characterization
45 Proprietary & Confidential
Dryspot Latex Reagents
• Completely dry including Control Reagents • less handling • no dispensing errors • Less chance of contamination
• Room temperature storage
• fridge space
• Extended shelf life • 2 years
• Improved reproducibility
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spotted Agglutinate with antigen Extract
5. 4.
Antibody absorbed to latex
Dry formulation spotted
Bacterial suspension or sera added
Agglutination observed
• Reagent Formulation
DrySpot Latex Reagents
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Features & benefits
Sample Collection
Sample Enrichment
/Dilution
Sample Prep/
Purification Detection Confirmation Identification/
Characterization
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Latex Agglutination Competitive Advantage
Sample Collection
Sample Enrichment
/Dilution
Sample Prep/
Purification Detection Confirmation Identification/
Characterization
• Large range which includes ‘’Gold Standard’’ kits
• Test and Control reagent eliminates reporting of non specific agglutination
• Wellcolex patented coloured latex technology
• DrySpot room temperature storage and extended shelf life
• One step in the end-to-end workflow solution offered by Thermo Scientific™
50 Proprietary & Confidential
Water Testing – laboratory workflow
51 Proprietary & Confidential
Legionnaire’s Disease
• Legionnaires’ disease, named after the outbreak in 1976 at the American Legion Convention in Philadelphia
• Caused by Legionella pneumophila and other Legionella species
• Characterised as an acute febrile respiratory illness ranging in severity from mild illness to fatal pneumonia
• The mortality rate can be as high as 25% in untreated immunosuppressed patients
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• Outbreaks of Legionnaires' disease receive significant media attention, especially when a large number of people become ill or die
• Single infections often go unnoticed. The Centers for Disease Control and Prevention (CDC) estimate between 8,000 and 18,000 people are hospitalized with Legionnaires' disease in the US each year.
• The major reservoir of Legionella appears to be fresh water sites, air-conditioning units and various water plumbing fixtures
• The immediate consequences for the building owner or manager faced with liability claims and negative publicity can be devastating and extremely costly.
• Proactively managing the risk of Legionella bacteria in cooling towers and water systems is more cost effective than responding to an outbreak
Legionnaire’s Disease
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Legionella
• Legionella pneumophila is the most common cause of Legionnaires’ disease.
• At present, 14 different serotypes exist of which Legionella pneumophila serogroup 1 accounts for 90% of cases
• Isolates derived from environmental and clinical samples may be cultured on standard non-selective or selective Legionella culture media.
• Legionella species on primary isolation have an absolute requirement for L-cysteine hydrochloride. Confirm that an isolate is a Legionella by showing that it cannot grow on any medium which does not contain L-cysteine hydrochloride.
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• Legionella CYE Agar Base (CM0655) • Charcoal Yeast Extract Agar for the isolation of Legionellaceae when used with
Legionella BCYE Growth Supplement SR0110 (Edelstein BYCE Agar)
• This medium can be further supplemented with one of the following: • Legionella BMPA Selective Supplement (SR0111) • Legionella GVPC Selective Supplement (SR0152) • Legionella GVPN Selective Supplement (SR0215) • Legionella MWY Selective Supplement (SR0118)
• Additionally, a medium omitting L-cysteine may be prepared from Legionella CYE Agar Base (CM0655) and BYCE Growth Supplement (SR0175)
Legionella Culture Media
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Oxoid Legionella Latex Test DR0800M • A fast and simple screening procedure for predominant pathogenic Legionella species and
serotypes
• The DR0800 (wet kit) kit contains: • DR0801 Legionella pneumophila serogroup 1 Test Reagent • DR0802 Legionella pneumophila serogroups 2–14 Test Reagent • DR0803 Legionella species Test Reagent • These 3 sub-kits are also available seperately
• The Legionella species reagent is reactive with the following species and serotypes: Legionella longbeachae 1 and 2; Legionella bozemanii 1 and 2; Legionella dumoffii; Legionella
gormanii; Legionella jordanis; Legionella micdadei; Legionella anisa
DrySpot format - same as the wet kit, only available as three separate kits: • DR0200 Legionella pneumophila serogroup 1 Test Reagent • DR0210 Legionella pneumophila serogroups 2–14 Test Reagent • DR0220 Legionella species Test Reagent
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• Recommended for the enumeration of enterococci in water supplies to detect and enumerate enterococci by the technique of membrane filtration or direct plating
• The medium is very selective for enterococci • Plates are incubated at 35oC for 4 hours and then at 44-45oC for 44 hours • All red or maroon colonies may be accepted as presumptive enterococci
Traditional Culture Media - Slanetz & Bartley Medium
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• ONPG – weak colour intensity and freely soluble, therefore unsuitable for use in solid media
First-use of chromogens
ONPG -ve ONPG +ve
• Demonstration of β-galactosidase production on tryptone nutrient agar
with ONPG discs
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E. coli
Chromophore component (e.g. indoxyl derivative such as 5-bromo-4-chloro-3-indoxyl)
Sugar component (e.g. galactose)
“X”-gal
Uptake
β-galactosidase
Chromogen
Background and Principles of Chromogenic Media
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Background and Principles of Chromogenic Media
NH
HO
5-bromo-4-chloro-3-indoxyl ('X')
Br
Cl• “X”-gal
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Background and Principles of Chromogenic Media
NH
HO
Cl
6-chloro-3-indoxyl ('Rose')
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• E. coli • E. coli O157 • Coliforms • Enterococci • VRE • Genera that cause urinary tract
infections • Staph. aureus • MRSA
Chromogenic Media Exist For….
• Listeria spp and L. monocytogenes
• Bacillus cereus • Enterobacter sakazakii • Clostridium perfringens • Vibrio parahaemolyticus • Candida spp, C. albicans
and C. tropicalis
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Culture Media Design
ß-galactosidase C8-esterase
α-glucosidase
ß-glucuronidase
ß-glucosidase
E.coli
Ent.sakazakii
Salmonella
Salmonella Shig. sonnei
Shig. sonnei
Shig. sonnei Shigella spp. Vibrio spp.
Pseudomonas spp.
Klebsiella spp. Enterobacter spp.
Citrobacter freundii
Citrobacter koseri
Sal. arizonae
Phenylalanine deaminase
Proteus spp. Providencia spp.
Hafnia alvei
Hafnia alvei
Vibrio vulnificus
P. vulgaris Enterobacter spp.
Serratia spp.
E.coli
Gram-negative organisms
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1. Growth of target organism
2. Inhibition of non-target organisms
3. Differentiation of target organisms
Three Key Principles of Design
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1. Growth of Target Organism Genus level or species level
• Careful selection of: • Amino acids and nitrogen nutrients • Peptones - acid or enzymatic digested protein rich extracts from Meat, Soya,
Casein & Pea or Yeast Extract Inclusion of specific vitamins and growth promoting factors
• Blood – e.g. Campylobacter • Serum • Vitamins Right energy source
• Carbohydrates – often glucose, but can be sucrose or other sugars Mineral salts and metals
• Common metals are Ca++,Mg++, Fe+++ • Trace metals can include Manganese, Zinc, Molybdenum and Copper
Key Principles of Design
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2. Inhibition of non-target organisms Antibiotics
• Often will inhibit G+ve or G-ve bacteria Chemical agents
• Inorganic salts - Bismuth, lithium, tellurite, tetrathionate Dyes
• Acriflavin, Brilliant green, crystal violet Surface active agents
• Bile, cetrimide, lauryl sulphate Others
• Dichloran, irgasan, Inhibigens
Key Principles of Design
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3. Differentiation of Target Organisms Chromogens
• Single or dual combination
pH indicator Dyes
• Phenol red .e.g Brilliant Green agar
Morphology
Biochemical
• Hydrogen sulphide e.g. XLD
Zone/halo formation due to proteolytic action
• e.g. Baird-Parker or Brilliance Listeria Natural colour of organism e.g. Staph aureus, Pseudomonas
Key Principles of Design
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• Formulation contains:
• Peptones and Yeast Extract – growth components
• Lactose – utilised by E.coli and coliforms producing acid
• Sodium Lauryl Sulphate – inhibits Gram +ve bacteria
• Phenol Red – pH sensitive dye used to colour colonies
• 5-bromo-4-chloro-3-indolyl-b -D-glucuronide – chromogenic substrate
• Agar – Gelling agent
Membrane Lactose Glucuronide Agar (MLGA)
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Differentiation of E.coli and coliforms is facilitated through two biochemical reactions:
1. Lactose fermentation by lowers the pH turning colonies yellow
2. Chromogenic substrate 5-bromo-4-chloro-3-indolyl-b-D-glucuronide (BCIG) is cleaved by the enzyme glucuronidase and produces a blue chromophore which builds up in the cells.
Membrane Lactose Glucuronide Agar – How Does It Work?
Organism Lactose fermentation
Glucuronidase enzyme
Colony colour
E.coli Yes (yellow) Yes (blue) Green coliforms Yes (yellow) No Yellow Pseudomonas No No Pink Bacillus (Gram +ve) - - Inhibited
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• Extensive product range
• Our core business is manual microbiology
• We are moving forward with technological advances
• BUT...... ......we must protect our market share and we must protect our customer base
in preparation for future developments
• There are still many opportunities , especially cross-selling products from the customer’s workflow
Our portfolio