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Evolution of MRSA During Hospital Transmission and Intercontinental Spread   (Harris  et al , 2010) Mariam Reyad Rizkallah 5 th  year student, Faculty of Pharmacy, Cairo University
[object Object],[object Object],[object Object],What is the problem? (Can’t depend on a snapshot) Genes that are transcribed at a relatively  constant level,  code for products for  maintaining the basic  cell  processes  or structure.
[object Object],[object Object],[object Object],[object Object],[object Object],What is the solution provided by the research?
Prospects ,[object Object]
Why  characterization  of pathogenic microorganisms? Characterization Local epidemiology: (hospital outbreaks)   Same or different  strain? Global epidemiology:   How  geographically-related?
What are the  current methods  used for the characterization of pathogenic microorganisms? ,[object Object],[object Object],[object Object],[object Object],[object Object]
Methods   The clone ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
[object Object],[object Object],Methods Genome-wide analysis, Genomic fingerprinting  & DNA comparison
Methods Genome-wide analysis, Genomic fingerprinting  & DNA comparison 63 isolates  2 samples 1 st  Sample 43 isolates Global collection (1982-2003) Intercontinental transmission 2 nd  Sample 20 isolates Northeast Thailand  (7-month period) Fine-scale transmission
Methods Genome-wide analysis, Genomic fingerprinting  & DNA comparison ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
Genome Analyzers ,[object Object],[object Object],[object Object],[object Object],Genome Analyzer IIe http://www.illumina.com/systems/genome_analyzer.ilmn
Genome Analyzer Applications http :// www . geneworks . com . au / library / MultipleApp_image . jpg
Genome Analyzer Applications ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
Results 6714 SNPs With marked  uneven distribution   across the genome Core  ( 4310) Chromosomal Vertical gene transfer Non-core MGEs Horizontal gene transfer (Major source)   38 homoplasic   genes involved in  drug resistance   “ similar in form & structure,  but not in origin”
Results ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
Results   –  Phylogenetic analysis & geographical relation
[object Object],[object Object],[object Object],[object Object],[object Object],Recent expansion of a single variant Single, but more diverse clade more diverse, most  positioned basally , consistent  Europe is the  possible origin for ST239 with geographical clustering
Source:  Pathogen Microevolution in High Resolution. (Aziz  et al , 2010,   Science Translational Medicine) A map showing the geographical spread of MRSA ST239 as deduced by Harris  et al .
[object Object],[object Object],[object Object],[object Object],[object Object],Portuguese clone,1990-1993 together  Explaining the relationship between ST239 clones in Portugal in the 1990s: Portuguese clone,1990s  Brazilian clone, 1997 Brazilian clone, 1997 clustered with South America isolates Second wave  in Portugal resulted from the  introduction of a South American variant
[object Object],[object Object],[object Object],[object Object],[object Object],phiSPb-like (TW20) prophage characteristic of the Asian clade. Single intercontinental transmission event from southeast Asia. Explaining the origin of London outbreak 2001-2003:  DEN907, isolate from Denmark, and TW20, from London among an  Asian clade ??
What did they find? Results ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
Results  -  Mutation rate   ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
Results  –   SNPs analysis   from  homoplasies in the core regions   SNP position Region/gene Isolate SNP (amino acid replaced, syn, intergenic) Antibiotic resistance Nucleotide replaced
Results  –   SNPs analysis   from  homoplasies in the core regions
Results  -  Fine - scale transmission ,[object Object],[object Object],[object Object],[object Object]
spa , PFGE vs. phylogenetic analysis
 
Prospects/Future applications ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
Comments & Questions
Thank you

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MRR_Harris_JC_20100217

  • 1. Evolution of MRSA During Hospital Transmission and Intercontinental Spread (Harris et al , 2010) Mariam Reyad Rizkallah 5 th year student, Faculty of Pharmacy, Cairo University
  • 2.
  • 3.
  • 4.
  • 5. Why characterization of pathogenic microorganisms? Characterization Local epidemiology: (hospital outbreaks) Same or different strain? Global epidemiology: How geographically-related?
  • 6.
  • 7.
  • 8.
  • 9. Methods Genome-wide analysis, Genomic fingerprinting & DNA comparison 63 isolates 2 samples 1 st Sample 43 isolates Global collection (1982-2003) Intercontinental transmission 2 nd Sample 20 isolates Northeast Thailand (7-month period) Fine-scale transmission
  • 10.
  • 11.
  • 12. Genome Analyzer Applications http :// www . geneworks . com . au / library / MultipleApp_image . jpg
  • 13.
  • 14. Results 6714 SNPs With marked uneven distribution across the genome Core ( 4310) Chromosomal Vertical gene transfer Non-core MGEs Horizontal gene transfer (Major source) 38 homoplasic genes involved in drug resistance “ similar in form & structure, but not in origin”
  • 15.
  • 16. Results – Phylogenetic analysis & geographical relation
  • 17.
  • 18. Source: Pathogen Microevolution in High Resolution. (Aziz et al , 2010, Science Translational Medicine) A map showing the geographical spread of MRSA ST239 as deduced by Harris et al .
  • 19.
  • 20.
  • 21.
  • 22.
  • 23. Results – SNPs analysis from homoplasies in the core regions SNP position Region/gene Isolate SNP (amino acid replaced, syn, intergenic) Antibiotic resistance Nucleotide replaced
  • 24. Results – SNPs analysis from homoplasies in the core regions
  • 25.
  • 26. spa , PFGE vs. phylogenetic analysis
  • 27.  
  • 28.