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Fourth Annual Digital PCR - 第4屆Digital PCR年度學會 -
2015年11月3 - 4日
美國,加州,聖地亞哥

Digital PCR學會:精密診斷的技術與工具– 第1天



聚合酶鏈鎖反應(PCR)對於核酸的檢測與定量是非常重要的工具。作為診斷實驗室不可或缺的要素,PCR隨著科學需求持續發展,成為最新的Digital PCR(dPCR)。dPCR不但提高了靈敏度與特異性,在許多情況下的通量也得到提升。Cambridge Healthtech Institute舉辦的第4屆Digital PCR年度學會,將聚焦對臨床診斷而言極重要的PCR進步。與往年一樣,將集結技術供應商及早期採用者、新平台參與者,在加深交流的同時針對Digital PCR的能力與限度、未來應用進行討論。本年度不僅聚焦dPCR,也將關注dPCR與NGS間界面、新興技術及研究領域。也將討論其他處理困難樣本的解決方案、提升樣本通量的方法,以及細胞外DNA研究、數位檢測指南及最佳範例等。

節目顧問

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


主要議題

  • 技術考量事項:晶片、微滴至多重化
  • DPCR驗證及參考標準
  • Digital PCR數據解析
  • dPCR與NGS的界面
  • 絕對定量
  • 複製數量變動
  • 少數標靶檢測

主要演講嘉賓


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

 

講習會*

Digital PCR:技術概要 

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

單細胞解析技術:準備狀況與能力 

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


*需另外報名參加。



11月3日(二)


第1天:DIGITAL PCR技術面向

7:30 am 登記報到與早晨茶敘

8:25 議長開會致辭

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

 

技術考量事項:晶片、微滴到多重化

8:30 結晶Digital PCR的導入

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之全基因組擴增

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 利用整合綜合微滴數位檢測快速檢測血液稀有生物標記

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 休息與展示會及海報發表欣賞


使用DPCR之驗證及參考標準

10:45 DNA標準物質的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 臨床評估上dPCR計數的潛在作用:以KRAS SNP為例

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 贊助商展演

12:15 pm 午餐展演或各自午餐

1:00 會議休息


DIGITAL PCR數據解析

1:25 議長致辭

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

 

1:30 廣義線性混合模型(GLMM)之微滴Digital PCR數據解析

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在微流體基質中in situ PCR之CRC組織内G12V KRAS檢測

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 贊助商展演

3:00 休息與展示會及海報發表欣賞


NGS與DPCR間的界面

3:30 藉NGS及Digital PCR從晚期腫瘤進行多次活體組織切片中的突變檢測

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 拷貝數變異: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準確定量之課題與挑戰

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 歡迎招待會及展示會與海報發表欣賞

6:00 第1天閉幕


* 活動內容有可能不事先告知作更動及調整。



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