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Digital PCR Technology Report from IPR 


Digital PCR Conference: Technologies and Tools for Precision Diagnostics

Conference Proceeding CD Now Available
  • Speaker Presentations
  • Poster Abstracts
  • and More!
2013 Archived Content

 

Digital PCR, at its core, is simply a single molecule counting method that quantitatively measures absolute DNA and eliminates the need for standard curves. However, it has already shown potential to be a disruptive technology in many areas of diagnostics. The Second Annual Digital PCR event will bring together industry visionaries and early adopters to discuss digital PCR's capabilities, limitations, and future applications. Researchers will examine applications in cancer biomarker and rare mutation detection, non-invasive fetal DNA analysis, and infectious disease quantification, particularly in HIV. Novel digital PCR devices from startups and academic labs will be showcased. Other topics to be addressed include digital PCR integration with existing technologies, solutions for processing difficult samples as well as increasing sample throughput, and guidelines and best practices for digital detection.

Scientific Advisory Board

Kerry R. Emslie, Ph.D., Bioanalysis Group Manager, National Measurement Institute, Australia

Jim Huggett, B.Sc. (Hons), Ph.D., Science Leader, Nucleic Acid Metrology, Molecular & Cell Biology, LGC

Ross Haynes, Biological Science Technician, Biochemical Science Division, National Institute of Standards and Technology

 

MONDAY, OCTOBER 7


PRE-CONFERENCE SHORT COURSE 

9:00 am - 12:00 pm

Digital PCR Experiment Design and Primer

Jim Huggett, B.Sc. (Hons), Ph.D., Science Leader, Nucleic Acid Metrology, Molecular & Cell Biology, LGC

 

The presentation will introduce the concept of digital PCR (dPCR) and explain how it compares to other molecular methodologies, citing both advantages and disadvantages. Included in the discussion will be the application of dPCR to perform minority target detection, absolute quantification and measurement of copy number variation as well as the analysis of RNA using reverse transcriptase dPCR. The discussion will also include some of the of the specific challenges associated with performing this technique.

About Jim Huggett 

Dr Huggett started working with quantitative molecular measurement at Cardiff University where he began his PhD in 1997 using the first Light Cycler qPCR instrument in the UK. Since then a substantial amount of his interest has been looking at molecular diagnostics and the standardisation of molecular methods. He has worked with institutes across Africa setting up and supporting a number of qPCR laboratories and advised the WHO on the use of molecular methods. Dr Huggett is also one of the original authors of the MIQE guidelines and has a expanding interest in molecule quantification. For the last two years he has worked at LGC, one of the UKs national measurement institutes, which had the first dedicated digital PCR instrument in the UK and has published a number of papers on dPCR analysis.

Bio-RadGrowing Droplet Digital PCR Applications: Multiplexing, Linkage Analysis and EvaGreen 

Dianna Maar, Ph.D., Senior Scientist, Applications, Bio-Rad Laboratories

Over the last two years the QX100 has enabled users to measure DNA and RNA targets without a standard curve and with a new level of precision and sensitivity. Currently, ddPCR is being recognized for its strength in detecting and measuring copy number variable genes, rare single-nucleotide polymorphisms, rare species and low-fold gene expression. Here I will discuss some of these current applications as well as the expanding applications for ddPCR™. Growing applications include micro RNAs, single-cell gene expression, gene linkage, multiplexing and EvaGreen applications.


GOING DIGITAL: WHAT CAN IT DO? 

1:25 pm Chairperson's Opening Remarks

Philipp Angenendt, Ph.D., CTO, Inostics GmbH


» KEYNOTE PRESENTATION

1:30 Digital PCR: A Paradigm Shift in Molecular Measurement

Jim Huggett, B.Sc. (Hons), Ph.D., Science Leader, Nucleic Acid Metrology, Molecular & Cell Biology, LGC

Digital PCR (dPCR) offers a unique ability to perform sensitive molecular measurement without the need for a standard curve. By converting the real-time PCR analogue signal to a binary output, measurement is simplified considerably. This also results in improved precision, facilitating the measurement of smaller differences, and the format leads to more sensitive detection of minority mutations. This presentation will discuss the benefits as well as disadvantages of dPCR, discuss the digital MIQE guidelines and offer a prediction of how dPCR may impact molecular measurement in the future.

2:00 What Validation is Required for Digital PCR Measurements?

Leonardo Pinheiro, Ph.D., Senior Scientist, Bioanalysis Group, National Measurement Institute, Australia

One of the major strengths of digital PCR is the direct counting approach which can provide precise, quantitative measurements without reference to a calibrant. However, the accuracy of data depends on successful amplification from each template molecule and, like real-time PCR, assay optimization is still critical. The requirements for validation of digital PCR assays will be discussed in relation to applications such as copy number variation, rare mutant detection and target quantification.

2:30 Technical Explanation and Theory of Digital PCR: What Can Go Wrong and Why?

Ross Haynes, Biological Science Technician, Biochemical Science Division, National Institute of Standards and Technology

This presentation will briefly go over how digital PCR works,take an intuitive look at Poisson statistics, and focus on possible measurement issues. These measurement issues illustrate that some prior knowledge of the material may be required to make good measurements. Understanding the details of how digital PCR works will help users intelligently design experiments, troubleshoot, and analyze results.

3:00 Refreshment Break with Exhibit and Poster Viewing

3:45 Digital PCR: A Historical Perspective

Alec Morley, Emeritus Professor, Haematology and Genetic Pathology, Flinders University

The first publication to use the term "digital PCR" appeared in 1999 but during the preceding decade there had been a series of publications which used the method either to study the properties of single molecules or to quantify single molecules. The rise and fall of these early studies will be reviewed

PodcastDigital PCR: Looking Back and Moving Forward with Alec Morley 

Bio-Rad4:15 Droplet Digital PCR Applications for Next-Generation Sequencing

Nicholas J. Heredia, Ph.D., Staff Scientist, Biological Sciences Group, Bio-Rad Laboratories Inc. 

NGS platforms are currently being used to quantify rare species targets in samples.  Heavy processing of the starting template in NGS workflows can result in biases of the representation of these targets.  Here we will discuss Droplet Digital PCR applications for cross validating NGS data with respect to rare species targets, as well as the excellent power of ddPCR quantification of NGS libraries for optimal sequencing performance.  

4:45 Welcome Reception in Exhibit Hall

5:15 Close of Day

 

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