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Fourth Annual Digital PCR - 제4회 디지털 PCR 컨퍼런스 -
2015년 11월 3 - 4일
미국 캘리포니아주, 샌디에고

디지털 PCR 컨퍼런스:정밀 진단을 위한 기술 및 툴 - 첫째날



중합효소 연쇄 반응(PCR)은 핵산의 검출와 정량 평가에서 매우 중요한 툴입니다. 진단 시험소에서의 주요 기술로 PCR은 과학적 요구와 함께 계속 발전하며 최신 디지털 PCR(dPCR)이 되고 있습니다. dPCR은 감도와 특이성뿐만 아니라 많은 경우, 처리 능력도 향상되었습니다. Cambridge Healthtech Institute가 주최하는 제4회 디지털 PCR 컨퍼런스에서는 임상 진단을 위한 중요 PCR의 진화에 초점을 맞춥니다. 예년처럼 기술 프로바이더 및 얼리 어답터, 신규 플랫폼 업체가 참가하여 교류를 확대하고, 디지털 PCR의 능력과 한계, 향후 응용에 대해 논의합니다. 올해는 dPCR에 관한 기술에 초점을 맞추며, dPCR과 NGS 간 인터페이스, 신규 기술 및 연구 분야에도 주목합니다. 기타 토픽으로 처리가 어려운 시료를 위한 솔루션, 시료 처리 능력을 향상시키기 위한 솔루션과 세포외 DNA 연구, 디지털 탐지를 위한 가이드라인과 베스트 프랙티스 등도 다룰 예정입니다.

프로그램 어드바이저

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


주요 의제

  • 기술적 고려사항:칩, 액적에서 다중화로의 이동
  • DPCR을 이용한 밸리데이션 및 참조 기준
  • 디지털 PCR 데이터 분석
  • dPCR과 NGS 인터페이스
  • 절대적 정량
  • 복제수 변동
  • 소수 표적 탐지

주요 연사


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

 

쇼트코스*

디지털 PCR:기술적 개요 

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

단세포 분석 기술:준비 상황과 능력 

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


*별도 등록 필요



11월 3일(화)


첫째날:디지털 PCR의 기술적 측면

7:30 am 참가 등록 및 모닝커피

8:25 의장 개회사

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

 

기술적 고려사항:칩, 액적에서 다중화로의 이동

8:30 결정 디지털 PCR의 도입

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 액적 디지털 MDA를 이용한 전게놈 증폭

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 통합된 종합적 액적 디지털 탐지를 이용한 혈중 희소 바이오마커의 신속 탐지

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 휴식시간과 전시회 및 포스터 발표 관람


DPCR을 이용한 밸리데이션 및 참조 기준

10:45 DNA 표준 물질을 위한 디지털 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 임상 평가에서 dPCR에 의한 계수의 잠재적 역할:KRAS SNP 사례

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 스폰서 후원 프레젠테이션

12:15 pm 런천 프레젠테이션 또는 개별 점심식사

1:00 세션 브레이크


디지털 PCR 데이터 분석

1:25 의장 인사

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

 

1:30 일반화된 선형 혼합 모델에 의한 액적 디지털 PCR의 데이터 분석

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 HHistoMosaic의 미세유체 매트릭스에서 in situ PCR에 의한 CRC 조직내 G12V KRAS 검출

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 스폰서 후원 프레젠테이션

3:00 휴식시간과 전시회 및 포스터 발표 관람


NGS와 DPCR 간 인터페이스

3:30 NGS 및 디지털 PCR에 의한 진행성 종양으로부터의 복수 생검 유형에서의 변이 탐지

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 복제수 변동:디지털 PCR, NanoString 및 차세대 시퀀싱

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 디지털 PCR에 의한 NGS 라이브러리의 정확한 정량 평가를 위한 문제와 전략

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 웰컴 리셉션과 전시회 및 포스터 발표 관람

6:00 첫째날 폐회


* 주최측 사정에 따라 사전 예고없이 프로그램이 변경될 수 있습니다.



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