Single-molecule enzyme-linked immunosorbent assay detects serum proteins at subfemtomolar concentrations

被引：1609

作者：

Rissin, David M. ^{[1
]}

Kan, Cheuk W. ^{[1
]}

Campbell, Todd G. ^{[1
]}

Howes, Stuart C. ^{[1
]}

Fournier, David R. ^{[1
]}

Song, Linan ^{[1
]}

Piech, Tomasz ^{[1
]}

Patel, Purvish P. ^{[1
]}

Chang, Lei ^{[1
]}

Rivnak, Andrew J. ^{[1
]}

Ferrell, Evan P. ^{[1
]}

Randall, Jeffrey D. ^{[1
]}

Provuncher, Gail K. ^{[1
]}

Walt, David R. ^{[2
]}

Duffy, David C. ^{[1
]}

机构：

[1] Quanterix Corp, Cambridge, MA USA

[2] Tufts Univ, Dept Chem, Medford, MA 02155 USA

来源：

NATURE BIOTECHNOLOGY | 2010年 / 28卷 / 06期

关键词：

PROSTATE-SPECIFIC ANTIGEN; ULTRASENSITIVE DETECTION; BIOCHEMICAL RECURRENCE; RADICAL PROSTATECTOMY; ALZHEIMERS-DISEASE; FEMTOLITER ARRAYS; DETECTION LIMITS; IMMUNO-PCR; SENSITIVITY; BIOMARKERS;

D O I：

10.1038/nbt.1641

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

The ability to detect single protein molecules(1,2) in blood could accelerate the discovery and use of more sensitive diagnostic biomarkers. To detect low-abundance proteins in blood, we captured them on microscopic beads decorated with specific antibodies and then labeled the immunocomplexes (one or zero labeled target protein molecules per bead) with an enzymatic reporter capable of generating a fluorescent product. After isolating the beads in 50-fl reaction chambers designed to hold only a single bead, we used fluorescence imaging to detect single protein molecules. Our single-molecule enzyme-linked immunosorbent assay (digital ELISA) approach detected as few as similar to 10-20 enzyme-labeled complexes in 100 mu l of sample (similar to 10(-19) M) and routinely allowed detection of clinically relevant proteins in serum at concentrations (<10(-15) M) much lower than conventional ELISA(3-5). Digital ELISA detected prostate-specific antigen (PSA) in sera from patients who had undergone radical prostatectomy at concentrations as low as 14 fg/ml (0.4 fM). (C) 2010 Nature America, Inc. All rights reserved.

引用

页码：595 / 599

页数：5

共 30 条

[1] Sensitivity by combination: immuno-PCR and related technologies [J].