rapid microbiology website
HOME NEWS EVENTS  
Alicyclobacillus
Bacillus
Campylobacter
Candida
Chlamydia
Clostridium difficile
CMV
Coliforms
Cryptosporidium
E. coli
E. coli O157
Enterobacteriacea
E. sakazakii
Enterococci
Enteric viruses
Giardia
Helicobacter
Hepatitis
HIV
Legionella
Listeria
MMR viruses
Molds [Moulds]
Mycobacterium
Neisseria
Pseudomonas
Resp. viruses
Salmonella
Shigella
Stapyhlococcus
Staph. aureus
Staph. [MRSA]
Streptococci
Toxoplasma
Yersinia
Yeasts
Vibrio


Manual Cultural Techniques for Rapid Microbiology
- there's life in them yet!


Key Characteristics
  • Techniques that rely on organism growth and metabolism to produce a characteristic reaction


  • most commonly used approach are media containing chromogens

  • easy to validate, low cost per test


Introduction
Microbiology is on a cusp; on one side, for almost twenty years the word rapid has been applied to developments in methodology reliant upon growth-based, cultural techniques. On the other side, the real-time polymerase chain reaction (RT-PCR) assays can now offer same day results with previously unimaginable accuracy and are poised to turn the microbiology lab into the molecular diagnostics department.

However these techniques normally come at an 'added value' price that many labs either cannot afford or cannot justify, if for example there is no definable cost benefit to the organisation in real time results. These labs require tests that are low capital and consumable cost, easy to establish and validate, low maintainance and need minimal technician training.

So there is still very definately a need for novel cultural technologies designed to take days off the standard time to result. Let’s examine what’s on offer for rapid manual cultural microbiology - this may be the last stop before PCRville but they still offer many advantages.
Technology
At a practical level, microbiology can be split between pathogen testing and quality testing, i.e. those organisms that will cause product spoilage and stability issues.  It is also possible to draw a line between true cultural techniques and those utilizing growth as the reporter but adapting the presentation of the media to include devices other than Petri dishes, and invariably an instrument of some description, here we are just going to focus on novel cultural media.

As a general rule, with the exception of fungal tests, most quality indicator organisms can be reported within 48 hours utilizing traditional media inoculated direct from a homogonized sample. These media formulations have been relatively unchanged for at least the last twenty-five years with efficacy largely empirically derived.

In terms of time to result, the same cannot be said for the pathogens. With the requirement for more sensitive detection limits, an enrichment stage, or stages, need to be included followed by confirmatory tests. This not only extends the time to result, (and this can be up to seven days for Salmonella spp and Listeria spp), but also increases labor and consumable costs. With commercial pressures acting to reduce the reporting timeframe, traditional growth media formulations began to evolve.

Leading this change away from traditional, empirically derived selective and diagnostic features were the chromogenic and fluorogenic plating media. Such media allowed species specific diagnostic features to be designed into direct plating media that would previously have required sub-culture and further testing. An early example would be the inclusion of the fluorogenic substrate methylumbelliferone -β -D-glucoronide (MUG) into bile salt media for the specific diagnosis of E.coli amongst lactose fermenting, bile tolerant Gram negative flora.

Chromogenic substrates followed closely, allowing colour to report the presence of specific species amongst closely related accompanying flora. Early examples were designed to report extra-cellular enzymatic activity and as such the reactions were not colony associated and diffusion of the colour into the surrounding media occurred. As with all emerging technologies there were some early problems and the regulatory framework was slow to adopt the novel media. However the take-home message for microbiologists was clear; cultural techniques could now be used to reduce extended reporting timescales by making species specific diagnostics available on primary plating media.

The latest generation of chromogenic media take this even further. From an improved understanding of genotype and gene expression, specific intra-cellular enzyme systems are targeted with substrates that produce colony associated colour reactions. Also, combining enzyme/substrate systems within the same medium allows differential diagnosis to take place, e.g. UTI media that can report the presence of 2 or 3 target groups of organisms on one plate.

The regulatory acceptance has also moved on with ISO standards now incorporating chromogenic media. The very latest innovation addresses the reduction of the time taken to enrich target pathogens in recovery systems designed to lead into chromogenic diagnostic media. By combining a toxin and a substrate, rather than a chromogen and a substrate it is now possible to specifically inhibit a competitor species within a recovery system. If that recovery system also contains specific growth promoters for the target species then the result is faster growth promotion allowing earlier inoculation onto specifically designed chromogenic plating medium.

Such systems designed for Salmonella spp and Listeria spp allow diagnostic recovery within 48 hours as opposed to a more typical 5 day test.  These cultural techniques have addressed the commercial pressures to report earlier on the microbiological status of valuable inventory.

 


Benefits

Chromogenic cultural techniques offer reporting timeframes more in line with commercial needs

Start-up costs are significantly lower than instrumental techniques

Improved regulatory recognition makes them more accessible for SME organizations.



Where do I get more information?
Suppliers by Sector:
News items:

Search rapidmicrobiology.com News Items:


Accreditation
Air Samplers
Antibiotic sensitivity
Atmosphere Systems
Autoclaves/Sterilizers
Auto media prep
Biodefense
Blood culture
Clean Rooms
Counting [TVC]
Endotoxin
Env.mon surfaces
Identification
Lab. automation
Lab. disposables
LIMS
Media Fill Trials
Media [chromogenic]
Media (dehydrated)
Media [prepared]
Pharma harmonization
Point of Care
QC Organisms
Rapid test (automated)
Rapid Method Culture
Ready to use Media
Sample Collection
Stains and Markers
Sterility testing
Swabs
Toxin tests
Urinary pathogens
Comments about this page?


Top |  Home |  Contact |  Site Map |  News |  Find Laboratory |  Labs. by Country |  Events
©2007 Copyright MicroLink