追蹤食品中的沙門氏菌和李斯特菌：快速檢測和菌株鑒別.pdf

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1、Rapid DetectionChasing Salmonella and Listeria in Food:Rapid detection and strain discriminationOlaf Degen,Director Industry MicrobioogyBruker 09 May 2024|2023 BrukerBRUKER MICROBIOLOGY&INFECTION DIAGNOSTICS2 2SalmonellaCronobacterColiformsCampylobacterProteomics Technology:MALDI-TOF MS-you can util

2、ize it forFood&Industrial Microbiology13/05/2024L.monocytogenesLactic acid bacteriaYeasts&MoldsFilamentous Fungiconfirmation of:identification of:Degen,Food Safety Summit 2024|2023 BrukerXLD AgarGRAM-NEGATIVE FOOD PATHOGENSDAY 1DAY 1Liquid enrichmentDAY 2DAY 2Transfer to selectiveagars:Streak on XLD

3、&Chromogenic Agar-Incubation:37.0C,24hDAY 3DAY 3Presumptive Salmonella,needs confirmationSAVE workload and Time-to-resultISO ISO 6579:Salmonella Salmonella ScreeningISO-confirmationConfirmation byMALDI BiotyperDAY 4&5DAY 4&5(normal workflow):Confirmation includes plating on non-selective Agar plates

4、 for 24h and biochemical tests.In many cases followed by Salmonella serology(subtyping).3 3Quality control of ingredientsEnd product checkA typical customer application:Confirmation of SalmonellaSamplingEnvironmental monitoringFood&Industrial Microbiology13/05/2024Degen,Food Safety Summit 2024|2023

5、BrukerMALDI-TOF MS FOR FOOD MICROBIOLOGYMALDI BiotyperprincipleSalmonellaListeriaIdentify Gram+,Gram-bacteria,yeasts and moldsin minutes,by their unique proteomic fingerprint Non-selectiveor selectiveagars4 413.05.2024Innovation with Integrity Food and Industrial MicrobiologyCronobacter|2023 Bruker|

6、2023 BrukerMALDI Biotyper ISO 16140 validated,plusAOAC-OMA method and method of USDA-FSIS laboratory guidebookMALDI BiotyperMALDI Biotyperhas been successfully tested by the has been successfully tested by the USDAUSDA-Food Safety and Inspection Service(FSIS).Food Safety and Inspection Service(FSIS)

7、.A flow chart specific for FSIS Laboratory Isolation and Identification of Salmonella is available(issued 2021):Test on Bruker MALDI Biotyper,starting from isolated colonies,allows final confirmationBrukers MALDI Biotypercan replace other identification technologies like VITEK2 Compact5 5Two AOAC OM

8、A methodsAOAC OMA#2017.09:Confirmation and Identification of Salmonella spp.,Cronobacter spp.,Campylobacter spp.,and other gram-negative organismsAOAC OMA#2017.10:Confirmation and Identification of Listeria monocytogenes and Listeria spp.,and other gram-positive organisms 13.05.2024Innovation with I

9、ntegrity Food and Industrial MicrobiologyFour MICROVAL certificatesSalmonellaListeria incl.L.monocytogenesCampylobacterCronobacterReports of the ISO 16140-part 6 validation studies are available.|2023 BrukerMALDI BIOTYPER BIOINFORMATICS13.05.20246Innovation with Integrity Food and Industrial Microbi

10、ology6Unknown microorganism is matched against each Main Spectrum in the libraryCalculation of a matching scoreEasy result presentation by traffic light“displayGreen=high-confidence species IDExport of raw data possibleMALDI BiotyperLibrary(RUO/GP)and our software:Easy result interpretation for food

11、 labsBruker MBT Compass Library(version 2023)4,274 different species 2,365 gram-positives 1,648 gram-negatives 220 yeasts 41 filamentous fungi|2023 BrukerMALDI BIOTYPERFOR FOOD INDUSTRYProduct contaminations are rapidly identified with the same workflowInIn-house workflowhouse workflowGrowth and cou

12、ntingCharacterization of the dominant bacteria/yeast/mold1.Yogurt can be contaminated e.g.,with Aspergillus flavus,A.parasiticus and A.nomius(Aflatoxin producer!)2.Some bacteria(Lactobacillus,Pediococcus,Pectinatus,and Megasphaera)or fungi(Fusarium)can generate off-flavors,turbidity and acidity in b

13、eerC Characterizationharacterization of the dominant bacteria/yeast/moldNo matter which product or microorganism type,the MALDI Biotyper enables rapid identification with one workflow for all.7 713.05.2024Innovation with Integrity Food and Industrial Microbiology|2023 BrukerBRUKER MICROBIOLOGY&INFEC

14、TION DIAGNOSTICSSource tracking and strain comparison needed?Salmonella O-serotypes(classifier)Dendrogram for clear visualization ofstrain discrimination forSalmonella O-groups and serotypesFood&Industrial Microbiology13/05/20248Degen,Food Safety Summit 2024|2023 Bruker|2023 BrukerMALDI BIOTYPER&IR

15、BIOTYPERMALDI Biotyper&IR Biotyperfor actionable insightsWhy the Duo?MALDI BiotyperQuestion:What is it?Question:What is it?Based on MALDIMALDI-MSMS and specific peptide/protein fingerprints Gold standard in fast Microorganism ID Microorganism ID on species level Large database Large database of 4600

16、 species The MBT Subtyping Module(RUO)allows some(sub)species differentiationIR BiotyperQuestion:Is there a difference between these Question:Is there a difference between these isolates?Or is it the same?isolates?Or is it the same?Based on“cell phenotyping”by FTFT-Infrared Infrared Spectroscopy(FTS

17、pectroscopy(FT-IR)IR)Strain typing method,Strain typing method,allows in-depth differentiation on isolate/strain level,below species(when the ID is known e.g.from MBT)“Real Time”Analysis:Used in Infection Control,“Real Time”Analysis:Used in Infection Control,analysis of transmission routes Classific

18、ationClassification based on machine learning(e.g.ANN)e.g.for serogrouping/subspecies determination9 9Food&Industrial Microbiology13/05/2024Degen,Food Safety Summit 2024|2023 BrukerBRUKER MICROBIOLOGY&INFECTION DIAGNOSTICSSource tracking and strain comparison needed?Salmonella O-serotypes(classifier

19、)E.Coli O157,O104:H4,more,customizedLocal discrimination possible,customized solutions(no classifiers)e.g.,for S.enterica serovar Typhimurium,S.enteritidis,S.gallinarum,S.kentuckyDendrogram for clear visualization ofstrain discrimination forSalmonella O-groups and serotypesFood&Industrial Microbiolo

20、gy13/05/202411Degen,Food Safety Summit 2024|2023 BrukerBRUKER MICROBIOLOGY&INFECTION DIAGNOSTICS1212Important gram-positive bacteriaListeria&Listeria monocytogenesFound in milk powder.fruits,nuts,meat,seafoodListeria in meat,seafood,vegetables,frozen foodFood&Industrial Microbiology13/05/2024Staphyl

21、ococcusStaphylococcus aureusaureusfood-borne pathogenDegen,Food Safety Summit 2024|2023 Bruker|2023 BrukerBRUKER MICROBIOLOGY&INFECTION DIAGNOSTICSSingle lab-validation for Listeria classifier in 20231313Food&Industrial Microbiology13/05/2024A validated IR Biotyper method(1)has beenintegrated into t

22、he accredited workflow for L.L.monocytogenesmonocytogenes analysisanalysisin food samplesaccording to ISO 11290:ISO 11290:MALDIMALDI-TOF MS TOF MS confirmationconfirmation on the speciesspecies levelsubsequent serogroupingserogrouping and pre-selection byFTIR FTIR spectroscopyspectroscopy for Whole

23、Genome Sequencing(WGS)(1)Source,In:Clinical Spectroscopy,Vol 5,December 2023:Validated differentiation of Listeria monocytogenes serogroups by FTIR spectroscopy using an Artificial Neural Network basedclassifier in an accredited official food control laboratory.https:/doi.org/10.1016/j.clispe.2023.1

24、00030 Degen,Food Safety Summit 2024|2023 Bruker|2023 BrukerIR BIOTYPER PUBLICATIONSPublication on Food-borne outbreak S.aureus 2023-June-2014Not for use in clinical diagnostic procedures.IR BiotyperSame clone!Food,Patient and Food,Patient and Food Handler isolatesFood Handler isolates IR Biotyper(co

25、nfirmed by WGS)was able to reveal the source of infection IR Biotyper is suitable for fastfast and low-cost epidemiological analysis of source attribution for S.aureus|2023 Bruker|2023 Bruker|MALDI BIOTYPER-EASY TO USE&PRECISE RESULTSBenefits of the combination MALDI Biotyper and IR Biotyper13.05.20

26、2415Innovation with Integrity Food and Industrial MicrobiologyMALDI-TOF MS technology based on profiling of proteins starting from colony material Very fast identification on species level One workflow for all microorganisms(bacteria,yeast,molds)Ease of use,only short training needed ISO 16140-6 cer

27、tified/AOAC-OMA/USDA-FSIS method for pathogens Annual library updates Own customer libraries possibleMBTIR BiotyperFT-IR-based strain comparison in light of phenotypic aspects starting from colony material Very fast strain discrimination Easy software for Cluster analysis by a variety of statistical

28、 methods Dedicated classifiers for targeted questions:Salmonella O-serogroups,Legionella pneumphila SG1|2023 Bruker|2023 Bruker Broadening the research applications for food authenticitytesting MBT allows e.g.,the analysis of meat or insect species Bruker is focusing on microbiology validations but

29、is supporting new applications from collaboration partners Publicly available collection of sample preparation protocols for MALDI-TOF MS based identification of meat,dairy products,fish and insects published and available from CVUAEXTENDED RESEARCH CAPABILITIESOutlook:MALDI Biotyperfor food fraud testing13.05.202416Dyk M et al.2020.Aspects of Food Control and Animal Health,Vol:13,Pages:1-13,ISSN:2196-3460.16Innovation with Integrity Food and Industrial MicrobiologyInnovation with Integrity Food and Industrial Microbiology