技术原理:
数字PCR( 也可称单分子PCR) 一般包括两部分内容,即PCR 扩增和荧光信号分析。在PCR 扩增阶段,与传统技术不同,数字PCR
一般需要将样品稀释到单分子水平,并平均分配到几十至几万个单元中进行反应。不同于qPCR 对每个循环进行实时荧光测定的方法,数字PCR
技术是在扩增结束后对每个反应单元的荧光信号进行采集。最后通过直接计数或泊松分布公式计算得到样品的原始浓度或含量。数字PCR
的定量方法不依赖于扩增曲线的循环阈值,因此不受扩增效率的影响,也不必采用看家基因和标准曲线,具有很好的准确度和重现性,可以实现绝对定量分析。在未来十年内,医学保健的目标是提供优质高效的个性化医疗。PCR
方法会在实现这个目标的过程中发挥重要作用。相比于传统的PCR,数字PCR 技术有着无可比拟的优势和广泛的应用前景。

数字PCR检测原理
应用领域:
·突变/稀有变异检测(Mutation/Rare
variant detection)
·拷贝数变异(Copy-number
variation)
·进入外周循环(包括血液和粪便)的肿瘤组织核酸分析
·无创产前筛查
·转化移植检测
·药理学(Pharmacogenetics)
·基因表达分析(Gene expression
analysis)-特别针对单细胞或少量细胞或者血清血浆样品
·线粒体拷贝数分析与线粒体突变分析
·数字PCR可以验证二代测序(NGS)
应用案例:
·Huggett
JF, Foy CA, Benes V et al.(2013) The digital MIQE guidelines: minimum
information for publication of quantitative digital PCR experiments. Clinical
Chem 6:892–902.
·Absolute
quantification by droplet digital PCRversus analog real-time PCR Christopher M
Hindson et al. nature methods | VOL.10 NO.10 | OCTOBER 2013.
·Detection
of Cancer DNA in Plasma of Early Stage Breast Cancer Patients Julia A. Beaver*1,
Danijela Jelovac February 19, 2014 American Association for Cancer Research.
·Evaluation
of a Droplet Digital Polymerase Chain Reaction Format for DNA Copy Number
Quantification Leonardo B. Pinheiro, Victoria A. Coleman,November 28, 2011
American Chemical.
·Qin
J, Jones R C, Ramakrishnan R. Studying copy number variations using a
nanofluidic platform. Nucleic Acids Res, 2008,36(18): e116.
·Lo
Y M D, Fiona M F, Chan K C A, et al. Digital PCR for the molecular detection of
fetal chromosomal aneuploidy. Proc Natl Acad Sci USA, 2007, 104(32):
13116-13121.
·Warren
L, David B, Irving L W, et al. Transcription factor profiling in individual
hematopoietic progenitors by digital RT-PCR. Proc Natl Acad Sci USA, 2006,
103(47): 17807-17812.
·Ottesen
E A, Jong W H, Stephen R Q, et al. Microfluidic digital PCR enables multigene
analysis of individual environmental bacteria. Science, 2006, 314(5804):
1464-1467.
·Tadmor
A D, Elizabeth A O, Jared R L, et al. Probing individual environmental bacteria
for viruses by using microfluidic digital PCR. Science, 2011, 333(6038): 58-62.
·Tian
Wang, Hongying Sha. , et al. Polar Body Genome Transfer for Preventing the
Transmission of Inherited Mitochondrial Diseases。Cell 157, 1591–1604, June 19,
2014.
![]() |
![]() |
|
![]() |
![]() |
![]() |
![]() |
![]() |
![]() |
![]() |
![]() |
![]() |