National Human Genome Research Institute Newborn Screening Study at NIH 2009 Cystinosis Research Network Family Conference...
 
Blood sample
Extract DNA Amplify DNA
Amplify the targeted DNA
Identify the targeted DNA
 
A treatment is available : Cysteamine <ul><li>Preserves renal function </li></ul><ul><li>Allows a normal growth rate </li...
<ul><li>Cystinosis is a genetic disease </li></ul><ul><li>The gene is known </li></ul><ul><li>Mutations are known </li></u...
 
Extraction and Amplification Cell lysate Genomiphi Genomeplex
DNA Mutation Primer Forward Primer Reverse Amplify the targeted DNA PCR Polymerase Chain Reaction
Amplify the targeted DNA PCR Polymerase Chain Reaction
Identify the targeted DNA <ul><li>Sequencing </li></ul><ul><li>Denaturing High Performance Liquid Chromatography (DHPLC) <...
A bead technology
Dye 1 Dye 2
Multiplexable Reaction
 
Example of results
Homozygote WT Homozygote Mut Heterozygote
National Human Genome Research Institute Conclusion Medical Genetics Branch
Acknowledgment Barbara C. Yang Tiffani Manolis Robert Moss Dr. David R. Adams Dr. Marjan Huizing Dr. William A. Gahl Dr. M...
Acknowledgment Barbara C. Yang Tiffani Manolis Robert Moss Dr. David R. Adams Dr. Marjan Huizing Dr. William A. Gahl Dr. M...
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National Human Genome Research Institute

Published on: Mar 3, 2016
Source: www.slideshare.net


Transcripts - National Human Genome Research Institute

  • 1. National Human Genome Research Institute Newborn Screening Study at NIH 2009 Cystinosis Research Network Family Conference Thierry Vilboux, Ph.D. National Institutes of Health National Human Genome Research Institute Medical Genetics Branch Human Biochemical Genetics Section Medical Genetics Branch
  • 3. Blood sample
  • 4. Extract DNA Amplify DNA
  • 5. Amplify the targeted DNA
  • 6. Identify the targeted DNA
  • 8. A treatment is available : Cysteamine <ul><li>Preserves renal function </li></ul><ul><li>Allows a normal growth rate </li></ul><ul><li>Preserves thyroid function </li></ul><ul><li>Depletes muscle & liver of cystine </li></ul>Early diagnosis and treatment are critical, this is why we pursued newborn screening for cystinosis.
  • 9. <ul><li>Cystinosis is a genetic disease </li></ul><ul><li>The gene is known </li></ul><ul><li>Mutations are known </li></ul>Disadvantages <ul><li>From a blood spot, small amount of DNA </li></ul><ul><li>90 mutations in CTNS identified </li></ul><ul><li>High heterogeneity type of mutations </li></ul><ul><li>Differentiate carriers from patients </li></ul><ul><li>Rapid and conducive to high throughput </li></ul>Advantages
  • 11. Extraction and Amplification Cell lysate Genomiphi Genomeplex
  • 12. DNA Mutation Primer Forward Primer Reverse Amplify the targeted DNA PCR Polymerase Chain Reaction
  • 13. Amplify the targeted DNA PCR Polymerase Chain Reaction
  • 14. Identify the targeted DNA <ul><li>Sequencing </li></ul><ul><li>Denaturing High Performance Liquid Chromatography (DHPLC) </li></ul><ul><li>Denaturing Gradient Gel Electrophoresis (DGGE) </li></ul><ul><li>Single strand Conformation Polymorphism (SSCP) </li></ul><ul><li>Multiplex Ligation-dependent Probe Amplification (MLPA) </li></ul><ul><li>XMAP (Luminex) </li></ul><ul><li>SNP chip </li></ul><ul><li>iPLEX (Sequenom) </li></ul><ul><li>Comparative Genomic Hybridization (CGH) </li></ul><ul><li>… </li></ul>
  • 15. A bead technology
  • 16. Dye 1 Dye 2
  • 17. Multiplexable Reaction
  • 19. Example of results
  • 20. Homozygote WT Homozygote Mut Heterozygote
  • 21. National Human Genome Research Institute Conclusion Medical Genetics Branch
  • 22. Acknowledgment Barbara C. Yang Tiffani Manolis Robert Moss Dr. David R. Adams Dr. Marjan Huizing Dr. William A. Gahl Dr. Michael R. Erdos
  • 23. Acknowledgment Barbara C. Yang Tiffani Manolis Robert Moss Dr. David R. Adams Dr. Marjan Huizing Dr. William A. Gahl Dr. Michael R. Erdos

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