The Phene PlateTM system Fingerprinting of bacteria in microplates PhPlate Microplate techniques

Principles of bacterial typing Genotyping - analysis of the bacterial genome Kemotyping - analysis of the contents of the bacterial cell or the cell envelope Phenotyping - analysis of the expression of the bacterial cell

Different kinds of bacterial fingerprints DNA fingerprint Biochemical fingerprint

The PhenePlateTM system can e.g. be used for Areas of application The PhenePlateTM system can e.g. be used for Microbial ecology Measures diversities of microfloras Measures stability of the microflora Measures similarity between different microfloras Epidemiology Can be used to verify whether isolates are of different clonal origin Can be used as a pre-screening method prior to DNA fingerprinting

How can you trace bacteria in surface water, drinking water, or food 1) Inoculate or 16 – 24 colonies of indicator bacteria per sample in PhenePlates™ 2) Read the result and analyze with the PhP-software

Tracing sources of bacterial pollutions in surface water Industrial effluents Human fecal material Animal waste

Tracing bacteria in consumers water Low diversity = Regrowth within the system High diversity = Inadequate purification

Tracing bacteria in food

How can you - simply detect a food borne outbreak caused by e. g How can you  - simply detect a food borne outbreak caused by e.g. ETEC or EHEC ?  1) Inoculate 8 or 16 E. coli colonies per sample (including food) in RE PhenePlates™ 2) Read the result and analyze with the PhP-software

…Food borne outbreak If no common strain – no outbreak ! 3) Look at the dendrogram for a common (outbreak) strain. If no common strain – no outbreak ! Common outbreak strain Food Patient 1 Patient 2 Patient 3 For confirmation, analyze only one ore a few isolates belonging to the common strain for appropriate properties

How can you - find out whether the same strain of Enterococcus sp How can you  - find out whether the same strain of Enterococcus sp. causes the UTI of Mrs. Svenson in ward A as in Mrs. Olsen in ward B ? Srain name PhP type SvensonA 1 SvensonB OlsenA OlsenB Control-1 2 Control-2 3 1.Inoculate the strains in the FS-PhenePlate™ 2.Read and analyze the result with the PhP-software 3.Look at the computer generated list of identical strains   PS: It works with Escherichia coli etc. as well

How can you - find out whether the Klebsiella strains in the neonatal ward originate from a common source ? 1) Inoculate the strains in the PhP-48 plates 2) Read and analyze the result with the PhP-software 3) Look at the dendrogram for a common outbreak strain  Common outbreak strain

How can I - reduce the number of isolates that need to be DNA typed? 1)Inoculate all isolates into the PhP plates ™ 2)Read and analyze the result with the PhP-software 3)Look at the dendrogram, and select only those isolates that could not be differentiated by PhP for DNA typing

How does PhP typing perform in comparison to DNA typing? Example: Clustered PhP data and PFGE patterns for VRE isolates

Checking bacterial cultures for purity Pick one pure colony and one pooled colony (or streak) for PhP typing Different biochemical fingerprints – low correlations - indicate unpure cultures Identical biochemical fingerprints – high correlations - indicate pure cultures D1 col r= 0.997 99 22 2 24 24 23 22 22 23 9 8 6 D1 pool 99 21 2 23 23 23 22 21 23 10 9 6   D3 col r= 0.907 99 22 8 8 9 13 21 21 24 23 10 14 D3 pool 99 21 5 8 8 13 21 22 21 14 7 14 D4 col r= 0.995 99 23 3 22 23 24 23 22 24 17 24 6 D4 pool 99 23 2 22 24 24 23 22 24 17 22 6 D6 col r= 0.706 99 23 2 23 6 23 23 22 24 12 8 6 D6 pool 99 24 5 9 8 8 23 22 24 11 10 8

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