Cambridge Healthtech Institute’s 6th Annual
Digital PCR
Technologies & Tools for Precision Diagnostics
August 18, 2017 | Grand Hyatt Washington | Washington, DC
Digital PCR remains an important technology to have in diagnostic labs, and as more groups add and use this technology, novel applications continue to emerge. Cambridge Healthtech Institute’s 6th Annual Digital PCR symposium will continue to address technological aspects of dPCR technologies, with a particular focus on integrating dPCR with other technologies like NGS. This symposium will also address applications of digital PCR, including cancer and infectious disease diagnostics. New this year will be a focus on how dPCR can be used to validate CRISPR experiments. Hear from researchers, test developers, and platform users on their experiences and best practices for using this valuable technology.
Final Agenda
Recommended Short Course(s)*
SC19: Digital PCR: Applications and Advances
*Separate registration required
FRIDAY, AUGUST 18
8:00 am Registration & Morning Coffee
8:25 Chairperson’s Opening Remarks
Raghu Mirmira, M.D., Ph.D., Lilly Professor of Pediatric Diabetes, Professor of Pediatrics, Medicine, Cellular & Integrative Physiology, Biochemistry & Molecular Biology; Director, Center for Diabetes and Metabolic Diseases; Director, Medical Scientist Training Program, Pediatrics, Indiana University
8:30 KEYNOTE PRESENTATION: Impact of Instrumentation and Reagent Selection on Results of Viral Load Testing by Digital PCR
Randall T. Hayden, M.D., Director, Clinical and Molecular Microbiology, Department of Pathology, St. Jude Children’s Research Hospital
Digital PCR holds promise as a quantitative method based on reaction partitioning and end-point amplification that may reduce result variability. This talk will focus on the increasing variety of available platforms, reagents and applications for digital PCR, and how choices among those may affect viral load measurements.
9:00 Development of Genomic DNA Standards for Measurement of Cancer Biomarkers Using dPCR
Kenneth D. Cole, Ph.D., Group Leader, Bioassay Methods, Biosystems and Biomaterials Division, National Institute of Standards and Technology
NIST is developing genomic reference material standards for gene copy measurements of HER2, MET and EGFR, to improve cancer diagnostics. We have developed a number of dPCR assays for reference genes that allow us to normalize the copy number measurement in the abnormal karyotypes of many cancer cells. NIST is also working on standards for the measurements of ctDNA, a challenge due to the low concentrations and degraded size.
9:30 ddPCR to Measure and Accommodate Pre-Analytical Variability in cfDNA Samples and Guide Downstream NGS
Muhammed Murtaza, Ph.D., MBBS, Assistant Professor and Co-Director, Center for Noninvasive Diagnostics, Translational Genomics Research Institute
10:00 Single Cell Analysis with the Naica System
Laura Cavé, Sales Application Specialist, Application & Sales, Stilla Technologies
The Naica System uniquely allows users to image droplets and their contents, both pre- and post amplification, as well as recover droplets for downstream analysis. We have taken advantage of these features to develop exciting new applications for single cell analysis in mammalian cells and bacteria
10:15 Sponsored Presentation (Opportunity Available)
10:30 Coffee Break with Poster Viewing
11:00 Using NGS and ddPCR for Mutation Tracking in ctDNA to Predict Early Relapse of Breast Cancer Patients
Isaac Garcia-Murillas, Ph.D., Senior Scientific Officer, Breast Cancer Now Research Centre, The Institute of Cancer Research
Analysis of ctDNA in plasma by NGS and ddPCR can be used to monitor for minimal residual disease and identify early breast cancer patients at high risk of relapse. Identification of these patients at high risk of relapse would allow tailoring of adjuvant therapy approaches.
11:30 The Application of Digital PCR in Synthetic Biology
Alexandra Whale, Ph.D., Senior Researcher, Molecular Biology, LGC
With the development of ever growing numbers of molecular applications in the field of synthetic biology, precision in validation and characterization approaches are becoming increasingly important. This presentation will focus on the use of digital PCR in the context of the synthetic biology field.
12:00 pm Single-Step Detection of Diverse CRISPR-Cas9 Gene Editing Events in vivo
Brett A. Kaufman, Ph.D., Associate Professor, Medicine, University of Pittsburgh
CRISPR-Cas9 is currently the most flexible means to initiate gene editing via DNA damage repair, creating targeted mutations by recombination-mediated editing or indel mutations by non-homologous end joining. For in vivo editing, multiple alleles can be formed in each animal, creating significant genetic complexity. To solve this, we have developed a fast, high-specificity locked nucleic acid probe with an internal reference assay (drop-off PCR) this is sensitive and quantitative for measuring the extent of CRISPR-Cas9 mediated editing in transgenic founder mice.
12:30 Sponsored Presentation (Opportunity Available)
1:00 Luncheon Presentation (Sponsorship Opportunity Available) or Enjoy Lunch on Your Own
1:30 Session Break
2:00 Chairperson’s Remarks
Alexandra Whale, Ph.D., Senior Researcher, Molecular Biology, LGC
2:05 Utility of Digital PCR in Diagnosis and Monitoring of Lymphoma
Rashmi Kanagal-Shamanna, M.D., Assistant Professor, Hematopathology, Molecular Pathologist, MD Anderson Cancer Center
2:35 ddPCR for Copy Number Variant Analysis in Clinical Molecular Diagnostics
Sami S. Amr, Ph.D., FACMG, Associate Lab Director, Laboratory for Molecular Medicine, Partners HealthCare Personalized Medicine, Partners HealthCare
ddPCR offers a cost-effective and fast platform to perform copy-number variant analysis as a primary or a confirmatory assay for molecular diagnostics across a variety of disease areas. We will look at how ddPCR can be used to identify and confirm copy number variants in targeted gene panels and how this platform can enhance disease panel testing that relies on next generation sequencing.
3:05 Refreshment Break with Poster Viewing
3:35 Digital PCR as a Platform for the Next Generation of Diabetes Biomarkers
Raghu Mirmira, M.D., Ph.D., Lilly Professor of Pediatric Diabetes, Professor of Pediatrics, Medicine, Cellular & Integrative Physiology, Biochemistry & Molecular Biology; Director, Center for Diabetes and Metabolic Diseases; Director, Medical Scientist Training Program, Pediatrics, Indiana University
The identification of subjects at-risk for diabetes would allow the institution of therapies to prevent or mitigate diabetes. Despite their diverse origins, both type 1 and type 2 diabetes share common defects in islet β cell function. This presentation will focus on digital PCR to identify nucleic acid biomarkers of β cell stress, and how use of these biomarkers could enable a new generation of clinical studies on diabetes prevention.
4:05 30 Minute Pathogen-Specific Phenotypic Antibiotic Susceptibility Test Directly from Clinical Samples Using Digital LAMP Quantification
Travis S. Schlappi, Ph.D. Candidate, California Institute of Technology
Rapid antimicrobial susceptibility testing (AST) would decrease misuse and overuse of antibiotics. The “holy grail” of AST is a phenotype-based test that can be performed within a doctor visit (30 min.). To achieve this requires determining a pathogen’s susceptibility after a short antibiotic exposure. We used digital PCR (dPCR) to precisely measure DNA replication in bacteria exposed to antibiotics, which shortened the required antibiotic exposure time to 15 min and allowed high resolution quantification of DNA replication on timescales faster than cell division. We then optimized the chemistries and performed the entire digital AST (dAST) workflow directly from clinical UTI samples in less than 30 min. This work lays a foundation for the development of a rapid, point-of-care AST that would improve patient outcomes and strengthen global antibiotic stewardship.
4:35 Close of Symposia Programs