Rapid detection and typing methods for Listeria in the food industry

Transcription

1 2016, Santiago, Chile Rapid detection and typing methods for Listeria in the food industry

2 Detection of Listeria Culture method Immunoassay Fluorescent in situ hybridization (FISH) Biosensors Almost every month DNA-based amplification method 2

3 8 Ecology of Listeria monocytogenes Gram negative Infectious type pattern Gram positive Resistant to-environment pattern 3

4 Listeria monocytogenes in food plant 4

5 Rapid for Detection Oh, my god! We have L.monocytogenes in our product! L.monocytogenes 5

6 Rapid for Detection Our manager order me to clean up the line immediately.. 6

7 Rapid for Detection Need at least 7days to clean up all parts.. Can`t wait 7days!! 7

8 Rapid for Detection Let me find out the contamination point by swab test.. 8

9 Rapid for Detection Wait for bacteria coming out after incubation 3 days to get the final results. 9

10 Rapid for Detection You, a ridiculous human being!! Get the result within few hours! 10

11 Rapid for Detection OK I `ve got A contamination point! Being rapid is more important than being overly accurate! 11

12 Rapid for Detection Hey, here! Splendid!!! NO problem,, It takes only few minutes! 12

13 The multiplex detection of food pathogens PCR PCR condition (Various annealing temperatures and multiplication cycles) 65 13

14 New species festivals in the past 7 years.. New Listeria species New Listeria species New Listeria species New Listeria species New NewListeria Listeria species species New species of 11 Listeria in total was added in the past 7 years 14

15 Proposal by Renato H. Orsi & Martin Wiedmann Listeria Other genera Characteristics and distribution of Listeria spp., including Listeria species newly described Bon since 2009 Kimura Appl Microbiol Biotechnol (2016) 100:

16 Pulspulsed field gel electrophoresis (PFGE) 16

17 MLST (1998) (Multi Locus Sequencing Typing) 17

18 MLVA (multiple locus variable number tandem repeat analysis) 18

19 MLST in NGS era MLST based on NGS Conventional MLST PCR amplification of 5 7 genes In silico MLST(5~7 genes) 2012 Whole genome(wg)mlst 2013 Core genome(cg)mlst 2014 Rapid development in the past 4 years!! 19 19

20 Typing methods of Listeria 20 20

21 Whole genome MLST (2013) MLST=5~7 genes Whole genome MLST (wg MLST) MLST using all genomic genes!! Dr.Martin C. J. Maiden Inventor of MLST (1998) Gene by gene analysis MLST revisited: the gene by gene approach to bacterial 2013 October ; 11(10):

22 Core genome MLST (2015) Whole genome MLST Core genome + MLST Core genome = The genes that all the strains have in common Accessory genome = The genes that only some strains have Ruppitsch et al., J Clin Microbiol 53: (2015) Pightling et al., BMC Microbiology 15:224 (2015) 22 22

23 Scalability Scalability Other outbreaks Specific outbreak SNP Common language Cg(core genome) MLST Unification of definition of core genome Bon Kimura Is Necessary Listeria monocytogenes 23

24 WGS Analysis Using JSpecies Tool (2016) Burall et al. Simple, easy, rapid! GATGATGATCAT GATG 2 GATG 1 ATGA 2 ATGA 1 TGAT 2 TGAT 1 GATC 1 ATCA 1 TCAT 1 Burall et al. PLOS ONE DOI: /journal.pone (2016) 24 24

25 The development of a world common database Listeria monocytogenes Global Microbial Identifier 25

26 Rapid detection and typing methods for Listeria in the food industry THANK YOU FOR YOUR ATTENTION! 26