Absolute quantification by droplet digital PCR versus analog real-time PCR

被引：1173

作者：

Hindson, Christopher M. ^{[1
]}

Chevillet, John R. ^{[2
]}

Briggs, Hilary A. ^{[2
]}

Gallichotte, Emily N. ^{[2
]}

Ruf, Ingrid K. ^{[2
]}

Hindson, Benjamin J. ^{[1
]}

Vessella, Robert L. ^{[3
]}

Tewari, Muneesh ^{[2
,4
,5
]}

机构：

[1] Bio Rad Labs Inc, Digital Biol Ctr, Pleasanton, CA USA

[2] Fred Hutchinson Canc Res Ctr, Div Human Biol, Seattle, WA 98104 USA

[3] Univ Washington, Dept Urol, Seattle, WA 98195 USA

[4] Fred Hutchinson Canc Res Ctr, Div Clin Res, Seattle, WA 98104 USA

[5] Fred Hutchinson Canc Res Ctr, Div Publ Hlth Sci, Seattle, WA 98104 USA

来源：

NATURE METHODS | 2013年 / 10卷 / 10期

基金：

美国国家卫生研究院;

关键词：

DNA COPY NUMBER; MICRORNAS; QUANTITATION; CANCER; SERUM; RNA;

D O I：

10.1038/nmeth.2633

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Nanoliter-sized droplet technology paired with digital PCR (ddPCR) holds promise for highly precise, absolute nucleic acid quantification. Our comparison of microRNA quantification by ddPCR and real-time PCRCR revealed greater precision (coefficients of variation decreased 37-86%) and improved day-to-day reproducibility (by a factor of seven) of ddPCR but with comparable sensitivity. When we applied ddPCR to serum microRNA biomarker analysis, this translated to superior diagnostic performance for identifying individuals with cancer.

引用

页码：1003 / +

页数：6

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