Advances in qPCR and dPCR Agenda

Registration

Coffee & Networking in Exhibition Hall

In-vivo Single Cell Transcript Analyses for Systems Modelling
Philip Day, Reader, The University of Manchester, United Kingdom

Single cell measurements of DNA, mRNA and protein for bcr-abl was measured in K562 cells to produce a steady state model of BCR.ABL protein abundance per mRNA molecule. Impact of heterogeneity and development of in-vivo transcript measurements will be discussed.

Advanced Molecular Tools for Proteome Analyses
Masood Kamali-Moghaddam, Associated Professor, Uppsala University, Sweden

This presentation will illustrate a set of molecular tools with very high sensitivity and specificity for detection and localization of proteins and their interactions. Furthermore, the application of the methods in the search for biomarkers will be discussed.

Lunch & Networking in Exhibition Hall

Poster Viewing Session

Evidence Based Guidelines for the Pre-analytical Phase of RNA Testing in Blood Samples
Francesca Malentacchi, Postdoctoral Fellow, University Of Florence, Italy

Within the EC funded project SPIDIA (www.spidia.eu), pan-European external quality assessment (EQA) programmes were implemented for blood RNA based tests. The results demonstrated that the use of blood collection tubes containing cellular RNA stabilizers allows a reliable gene expression analysis whereas RNA measurements deriving from EDTA blood collection tubes showed an ex-vivo gene-dependent induction even 24h after blood collection. The results of the SPIDIA RNA EQAs studies have been used for the development of a Technical Specification by the European Committee for standardization (CEN).

qPCR in the Eyes of Europe’s No. 1. Commercial Service Supplier; Challenges and Innovative Solutions
Jens Bjorkman, CSO Manager, TATAA Biocenter AB, Sweden

Since the publication of the MIQE (Minimum Information for Publication of Quantitative Real-Time PCR Experiments) guidelines in 2009 users’ awareness and clients’ requests of quality control (QC) in qPCR has increased. I will describe some of our research into this field and provide real-life examples on how quality assurance protocols and strategies in qPCR workflow improve the performance and reliability of results.

Coffee & Networking in Exhibition Hall

Determination and Follow-up of a Mutational Profile with Circulating Tumour DNA Analysis
Florent Mouliere, Researcher, Nitzan Rosenfeld Group, University of Cambridge, United Kingdom

Circulating tumour DNA (ctDNA) exhibits the same genetic alterations from its tumour of origin, and can be used as a “liquid biopsy” for monitoring tumour progression. Q-PCR, digital PCR and next-generation sequencing are technologies of choice for determining and following-up a mutational profile with ctDNA.

Selective Amplification of Rare Mutant Sequences in Samples Containing Abundant Wild-type Sequences
Fred Kramer, Program Director, University of Medicine and Dentistry of New Jersey, United States of America

SuperSelective primers, by virtue of their unique design, employ thermodynamic and kinetic principles that enable the routine quantitation of as few as 10 mutant molecules in samples containing 1,000,000 wild-type molecules, even if the mutation is only a single-nucleotide polymorphism.

End of Day One

Finding Needles in a Haystack with Q-PCR – Rare Mutations, Single Cell Genotyping and DNA Methylation Analysis
Jorg Tost, Director, Centre National de Genotypage CEA, France

E-ice-COLD PCR enables reliable detection and sequence based identification of low prevalence mutations with clinical relevance in multiple types of patient samples including plasma. In addition, a simple protocol using standard real-time thermocyclers allows analyzing genotypes and DNA methylation patterns in cell-sorted single cells.

Coffee & Networking in Exhibition Hall

Digital PCR Goes COLD: Application of COLD-PCR in Digital Format Enables Quantitative and Multiplexed Mutation Scanning
Elena Castellanos-Rizaldos, Instructor in Radiation Oncology, Dana-Farber Cancer Institute/Harvard Medical School, United States of America

As currently applied, digital PCR can only be used to detect known mutations at single sequence positions. By merging COLD-PCR and digital -PCR technologies we enable digital mutation scanning, a new frontier for digital-PCR. A single reaction can now interrogate numerous mutations, and replaces multiple individual digital-PCR reactions.

Lunch & Networking in Exhibition Hall

Poster Viewing Session

Digital PCR in the Clinical Virology Laboratory - Applications and Limitations
Keith Jerome, Professor, University of Washington, United States of America

Digital PCR (dPCR) allows sensitive and accurate absolute quantitation of nucleic acid samples without the need for a standard curve. While dPCR is still mainly used in research settings, the characteristics of dPCR also make it attractive in the clinical laboratory. In this talk we will discuss applications of dPCR in the diagnostic virology laboratory, including ultraprecise cytomegalovirus monitoring, diagnosis of chromosomally integrated human herpesvirus 6, and quantitation of latent HIV reservoirs.

Quantification of Bacterial DNA for Metagenomic Control Materials
Denise O'Sullivan, Researcher, LGC Genomics, United Kingdom

In this study we use digital PCR to quantify the component organisms of a metagenomic control material comprising 10 common human pathogens mixed together in different proportions.

Coffee & Networking in Exhibition Hall

qPCR as a Tool for Early Detection of Alzheimer´s Disease
Petar Podlesniy, Postdoctoral Research Associate, Consejo Superior De Investigaciones Científicas, Spain

I will share our experience in the implementation of ddPCR for quantitation of low abundance targets (in the range of hundredths molecules per sample) as the circulating nucleic acids in biological fluids. Special emphasis will be made on the sample preparation and the controls used.

Analysing Circulating Tumor DNA in Clinical Trials – Today and in Future
Arndt Schmitz, Head/Senior Scientist, Bayer Schering Pharma AG, Germany

BAYER experiences with analysing 2,000 samples in our clinical trials with the most sensitive technique available will be reported. The field will soon be more complex, with established techniques becoming companion diagnostics and new technologies such as NGS entering.

Close of Conference