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Fourth Annual Digital PCR - 第4届Digital PCR年度学会 -
2015年11月3 - 4日
美国,加州,圣地亚哥

Digital PCR学会:精密诊断的技术与工具― 第1天



聚合酶链锁反应(PCR)对于核酸的检测与定量是非常重要的工具。作为诊断实验室不可或缺的要素,PCR随著科学需求持续发展,成为最新的Digital PCR(dPCR)。dPCR不但提高了灵敏度与特异性,在许多情况下的通量也得到提升。Cambridge Healthtech Institute举办的第4届Digital PCR年度学会,将聚焦对临床诊断而言极重要的PCR进步。与往年一样,将集结技术供应商及早期采用者、新平台参与者,在加深交流的同时针对Digital PCR的能力与限度、未来应用进行讨论。本年度不仅聚焦dPCR,也将关注dPCR与NGS间界面、新兴技术及研究领域。也将讨论其他处理困难样本的解决方案、提升样本通量的方法,以及细胞外DNA研究、数位检测指南及最佳范例等。

节目顾问

Jim Huggett, B.Sc. (Hons), Ph.D., Science Leader, Nucleic Acid Metrology, Molecular & Cell Biology, LGC
N. Somanath Bhat, Ph.D., Acting Senior Research Scientist, Bioanalysis Group, Department of Industry and Science, National Measurement Institute, Australia


主要议题

  • 技术考量事项:芯片、微滴至多重化
  • DPCR验证及参考标准
  • Digital PCR数据解析
  • dPCR与NGS的界面
  • 绝对定量
  • 复制数量变动
  • 少数标靶检测

主要演讲嘉宾


Jim HuggettJim Huggett, BSc (Hons), Ph.D., Science Leader, Nucleic Acid Metrology, Molecular & Cell Biology, LGC GroupFilip

 

Filip JankuFilip Janku, M.D., Ph.D., Assistant Professor, Investigational Cancer Therapeutics (Phase I Program), MD Anderson Cancer Center

 

Valerie TalyValerie Taly, Ph.D., Group Leader, CNRS Researcher, UMR S1147, University of Paris Descartes, CNRS

 

讲习会*

Digital PCR:技术概要 

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

单细胞解析技术:准备状况与能力 

Michael Masterman-Smith, Ph.D., CEO, Harmony Biosciences, Inc.


*需另外报名参加。



11月3日(二)


第1天:DIGITAL PCR技术面向

7:30 am 登记报到与早晨茶叙

8:25 议长开会致辞

N. Somanath Bhat, Ph.D., Acting Senior Research Scientist, Bioanalysis Group, Department of Industry and Science, National Measurement Institute 

 

技术考量事项:芯片、微滴到多重化

8:30 结晶Digital PCR的导入

Dangla RemiRémi Dangla, Ph.D., President and Co-Founder, Stilla Technologies

Stilla Technologies unveils its unique tool for high precision genetic analysis: Crystal digital PCR. Taking advantage of groundbreaking microfluidic advances, Crystal digital PCR relies on a single consumable to perform on-chip PCR in monolayer droplet arrays. This innovative approach increases multiplexing capacities in digital PCR and is readily amenable to large-scale, high-throughput clinical studies, thanks to a simple and fast workflow.

9:00 以微滴式Digital MDA之全基因组扩增

Minsoung RheeMinsoung Rhee, Ph.D., Postdoctoral Research Fellow, Biological Science and Technology, Sandia National Labs

We demonstrated for the first time that whole genome amplification by MDA can be performed in picoliter emulsion droplets, resulting in improved performance over corresponding reactions carried out at conventional microliter scale. De novo assembly of E. coli genome sequences from our droplet MDA and conventional tube MDA has revealed that droplet MDA showed more uniform coverage for all length of the genome and ~>99% of specific amplification.

9:30 利用集成综合微滴数位检测快速检测血液稀有生物标记

Dong-Ku KangDong-Ku Kang, Ph.D., Assistant Research Scientist, Sue and Bill Gross Stem Cell Research Center, Pharmaceutical Sciences & Biomedical Engineering, University of California, Irvine; Co-Founder and CSO, VeloxBiosystems

Rapid and sensitive diagnosis remains a major unmet challenge in food industry and medical applications. In this presentation, I will discuss a new technology called "Integrated Comprehensive Droplet Digital Detection Technology" (IC 3D) that is able to rapidly (1-2 h) and selectively detect rare pathogens/or rare biomarkers from milliliters (mLs) of complex media (e.g., unprocessed whole blood) at single-cell sensitivity in a one-step, homogenous, and culture-free reaction. This platform technology also has the potential to introduce a new paradigm in rapid detection for various diseases including cancer (targeting circulating tumor cells), neurological disorder (e.g., Alzheimer's disease), and infectious diseases (e.g., Lyme disease, HIV, and Ebola).

10:00 休息与展示会及海报发表欣赏


使用DPCR之验证及参考标准

10:45 DNA标准物质的Digital PCR

Somanath BhatN. Somanath Bhat, Ph.D., Acting Senior Research Scientist, Bioanalysis Group, Department of Industry and Science, National Measurement Institute

Accurate, reliable and reproducible quantification of nucleic acids is vital for many diagnostic applications and in routine laboratory testing. Digital PCR has the potential to not only improve quantitative nucleic acid analysis, but also to be considered as a reference method for certification of nucleic acid reference materials (RMs). This presentation will discuss how this technology was applied to characterize DNA RMs and discuss factors affecting reliability of the results.

11:15 临床评估上dPCR计数的潜在作用:以KRAS SNP为例

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

Unlike qPCR, dPCR does not require a calibration curve for quantitative analysis as the partitioning required to perform the technique enables DNA to be directly counted, or enumerated. This characteristic is fairly unique and opens a number of possibilities when considering clinical measurement, either through direct measurement using dPCR or in its use to support other methods, like qPCR, through the quantification of reference materials. This talk will discuss the work of the European Metrology Research Programme funded project Bio SITrace (http://biositrace.lgcgroup.com/) which is investigating the accuracy of dPCR when measuring rare single nucleotide polymorphisms in cell free DNA.

11:45 赞助商展演

12:15 pm 午餐展演或各自午餐

1:00 会议休息


DIGITAL PCR数据解析

1:25 议长致辞

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

 

1:30 广义线性混合模型(GLMM)之微滴Digital PCR数据解析

Oliver ThasOliver Thas, Ph.D., Professor, Biostatistics, Ghent University (Belgium) and University of Wollongong (Australia)

Target quantification with ddPCR depends heavily on the Poisson assumption. We demonstrate how Generalized Linear Mixed Models (GLMM) can be used for the data-analysis. GLMM is very flexible and allows for analyzing many designs, including dealing with replicates, run or plate effects and one or more references. The method can be used for absolute quantification, CNV and gene expression, and it allows for standard error and confidence interval calculation and hypothesis testing.

2:00 HHistoMosaic在微流体基质中in situ PCR之CRC组织内G12V KRAS检测

Emil KartalovEmil Kartalov, Ph.D., Assistant Professor, Pathology, Keck School of Medicine, University of Southern California

We present the experimental proof of principle of HistoMosaic - a novel technique for in situ genetic analysis of cancer tissue sections. HistoMosaic offers a high-throughput, low-cost, high-sensitivity means of detection and localization of rare mutations conferring drug resistance to cancer tissue, while the morphological information is preserved and co-registered with the genetic information. This ability would allow proper stratification of cancer patients, so the right drug is given to the right patient. HistoMosaic also has wide applicability in fundamental research and drug development.

2:30 赞助商展演

3:00 休息与展示会及海报发表欣赏


NGS与DPCR间的界面

3:30 藉NGS及Digital PCR从晚期肿瘤进行多次活体组织切片中的突变检测

Errin L. LagowErrin L. Lagow, Ph.D., Senior Scientific Liaison, Asuragen, Inc.

Targeted NGS permits detection of low-frequency somatic mutations in tumor biopsies, but call confidence may require independent assessment. Multiple biopsy types from advanced tumors were analyzed with the Quantidex™ PanCancer Panel, designed, developed, and cGMP-manufactured by Asuragen, and with digital PCR. Low frequency mutations were identified in FFPE tissue and confirmed in matched frozen tissue. Digital PCR demonstrated high utility for resolving discordant calls for samples with low frequency mutations.

4:00 拷贝数变异:Digital PCR、NanoString及次世代定序

Reinhold PollnerReinhold Pollner, Ph.D., Director, Clinical Trial Assay Development, Genoptix, Inc., a Novartis company

A variety of different technologies can be utilized to determine copy number variations in clinical samples. My presentation will focus on how three digital technologies - digital PCR, NanoString and Next-Generation Sequencing are used to determine copy number variations in clinical trial samples for patient stratification or exploratory purposes.

4:30 Digital PCR的NGS Library准确定量之课题与挑战

Peter SchweitzerPeter Schweitzer, Ph.D., Director, Genomics Facility, Institute of Biotechnology, Cornell University

One critical step in producing high quality DNA sequence data in a timely and cost efficient manner is accurately quantifying Illumina sequencing libraries. Digital PCR (dPCR) offers several advantages over real-time PCR. One significant challenge is accurately performing the large dilution required and I'll describe an internal reference developed to correct for variability during dilution. I'll also describe our experiences with dPCR assays for Illumina libraries using several different platforms.

5:00 欢迎招待会及展示会与海报发表欣赏

6:00 第1天闭幕


* 活动内容有可能不事先告知作更动及调整。



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