WEDNESDAY, NOVEMBER 4
8:00 am Morning Coffee
8:25 Chairperson’s Remarks
Stephen Bustin, Ph.D., FSB, Professor, Molecular Medicine, Postgraduate Medical Institute, Faculty of Medical Sciences, Anglia Ruskin University
8:30 Applications of Digital PCR in Clinical Microbiology
Randall T. Hayden, M.D., Director, Clinical and Molecular Microbiology, Member, Pathology, St. Jude Children’s Research Hospital
The use of digital PCR for clinical diagnostic applications will be reviewed, with a focus primarily (but not exclusively) on applications related to viral load testing. Discussion will include a comparison to real–time methods, and a review of current literature.
9:00 Digital PCR Quantification of Proteins
Stephen Bustin, Ph.D., FSB, Professor, Molecular Medicine, Postgraduate Medical Institute, Faculty of Medical Sciences, Anglia Ruskin University
The detection of proteins expressed by fungal and bacterial pathogens can be more clinically relevant than detection of their nucleic acids. Digital PCR can provide a quantitative readout of immuno-PCR-based methods, allowing precise and absolute quantification of pathogen load and making it a useful method for the early diagnosis of diseases where ELISAs are not sufficiently sensitive and PCR not adequately informative.
9:30 Rare Mutation Analysis Using Digital PCR Technology
Edward J Enright, M.S., Senior Technical Specialist, Thermo Fisher Scientific
10:00 Coffee Break with Exhibit and Poster Viewing
10:45 Molecular Analysis of Liquid Biopsies in the Diagnosis and Therapy of Advanced Cancers
Filip Janku, M.D., Ph.D., Assistant Professor, Investigational Cancer Therapeutics (Phase I Program), MD Anderson Cancer Center
The discovery of mutated “cancer genes,” has provided key insights into the mechanisms of tumorigenesis, which have been useful for developing targeted cancer therapies directed at specific druggable targets. Oncogenic mutations are tested using archival tumor tissue and its lack of availability is often a limiting factor. In addition, mutation analysis of primary tumor tissue or of an isolated metastasis does not necessarily reflect the genetic make-up of metastatic disease due to tumor heterogeneity. Liquid biopsies utilize cell-free DNA released to the circulation from cancer lesions or the tumor microenvironment. Molecular testing of cell-free DNA can provide valuable information about genetic changes that occur during the disease trajectory, as this is not a static process.
11:15 Tracking Circulating Tumor DNA Using Picoliter Droplet-Based Digital PCR
Valerie Taly, Ph.D., Group Leader, CNRS Researcher, UMR S1147, University of Paris Descartes, CNRS
Droplet-based digital PCR has recently emerged as a tremendous tool for cancer research and cancer patient follow-up. This presentation will illustrate how, by allowing non-invasive highly sensitive and quantitative analysis of circulating tumor DNA within blood patient samples, picoliter droplet-based PCR has been used for follow-up of cancer progression and tumor response to treatment.
11:45 Novel Approaches for Comprehensive Analyses of Circulating Cell-Free Tumor DNA
Hatim Husain, M.D., Assistant Professor of Medicine, Division of Hematology-Oncology, University of California, San Diego
The concept of liquid biopsies is expanding to include blood, urine, and other body fluids as specimen types. This talk will focus on the clinical translation of emerging technologies into treatment decisions and clinical trials to enabling development of novel algorithms to improve patient care.
12:15 pm Enjoy Lunch on Your Own
1:55 Chairperson’s Remarks
Hatim Husain, M.D., Assistant Professor of Medicine, Division of Hematology-Oncology, University of California, San Diego
2:00 Improving Plant Pathogen Diagnostics with Digital PCR
Tanja Dreo, Head, Bacteriology R&D, Biotechnology and Systems Biology, National Institute of Biology
The droplet digital PCR (dPCR) using QX100 (Bio-Rad) was assessed for detection and absolute quantification of quarantine plant pathogenic bacteria Erwinia amylovora and Ralstonia solanacearum. Different threshold settings were compared enabling selecting the approach leading to optimal diagnostic specificity on a set of defined control samples. Performance parameters were determined on a defined set of samples demonstrating equal or improved detection of both bacteria with digital PCR.
2:30 Multiplexing Strategies for Digital PCR in Karyotyping Yeast
William McDowell, Research Technician, Molecular Biology Facility, Stowers Institute for Medical Research
Multiplex reactions are an attractive option to increase the throughput and decrease the cost of digital PCR. We developed tetraplex assays for yeast karyotyping and discovered that cluster separation and discrimination of whole chromosome copy numbers were greatly affected by the sample preparation method and final assay concentrations. Best results were obtained with high quality genomic DNA templates and by limiting assay competition through optimization of assay and template concentrations.
3:00 Close of Conference
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