Digital droplet ELOHA for nucleic acid molecule counting and analysis

Weihua Guan; Liben Chen; Tushar Rane; Aniruddha Kaushik; Jeff Tza Huei Wang

doi:10.1109/TRANSDUCERS.2015.7180960

Digital droplet ELOHA for nucleic acid molecule counting and analysis

Weihua Guan, Liben Chen, Tushar Rane, Aniruddha Kaushik, Jeff Tza Huei Wang

Whiting School of Engineering

Research output: Chapter in Book/Report/Conference proceeding › Conference contribution

Abstract

We present a droplet digital Enzyme-Linked Oligonucleotide Hybridization Assay (droplet digital ELOHA) for reference-free absolute quantification of nucleic acids. Droplet digital ELOHA is able to accurately quantify as few as several hundred copies of nucleic acids in a large, mL-sized sample volume (equivalent to 1 aM or lower) without molecular replication. Droplet digital ELOHA provides a linear response to the number of copies present in a sample to allow for detecting a small fold change unattainable by real time quantitative PCR. Its capability to cope with a large sample volume benefits from the use of magnetic beads to streamline capturing of nucleic acids from the sample and subsequent surface hybridization reactions in droplets. Droplet digital ELOHA is particularly attractive for RNA quantification since it does not require reverse transcription. The absolute quantification ability of droplet digital ELOHA is demonstrated with the analysis of clinical Neisseria Gonorrhoeae 16S rRNA to show its potential diagnostic value.

Original language	English (US)
Title of host publication	2015 Transducers - 2015 18th International Conference on Solid-State Sensors, Actuators and Microsystems, TRANSDUCERS 2015
Publisher	Institute of Electrical and Electronics Engineers Inc.
Pages	460-463
Number of pages	4
ISBN (Electronic)	9781479989553
DOIs	https://doi.org/10.1109/TRANSDUCERS.2015.7180960
State	Published - Aug 5 2015
Event	18th International Conference on Solid-State Sensors, Actuators and Microsystems, TRANSDUCERS 2015 - Anchorage, United States Duration: Jun 21 2015 → Jun 25 2015

Publication series

Name	2015 Transducers - 2015 18th International Conference on Solid-State Sensors, Actuators and Microsystems, TRANSDUCERS 2015

Other

Other	18th International Conference on Solid-State Sensors, Actuators and Microsystems, TRANSDUCERS 2015
Country/Territory	United States
City	Anchorage
Period	6/21/15 → 6/25/15

Keywords

digital assay
Droplet microfluidics
single molecule counting

ASJC Scopus subject areas

Instrumentation
Electrical and Electronic Engineering

Access to Document

10.1109/TRANSDUCERS.2015.7180960

Cite this

Guan, W., Chen, L., Rane, T., Kaushik, A., & Wang, J. T. H. (2015). Digital droplet ELOHA for nucleic acid molecule counting and analysis. In 2015 Transducers - 2015 18th International Conference on Solid-State Sensors, Actuators and Microsystems, TRANSDUCERS 2015 (pp. 460-463). Article 7180960 (2015 Transducers - 2015 18th International Conference on Solid-State Sensors, Actuators and Microsystems, TRANSDUCERS 2015). Institute of Electrical and Electronics Engineers Inc.. https://doi.org/10.1109/TRANSDUCERS.2015.7180960

Digital droplet ELOHA for nucleic acid molecule counting and analysis. / Guan, Weihua; Chen, Liben; Rane, Tushar et al.
2015 Transducers - 2015 18th International Conference on Solid-State Sensors, Actuators and Microsystems, TRANSDUCERS 2015. Institute of Electrical and Electronics Engineers Inc., 2015. p. 460-463 7180960 (2015 Transducers - 2015 18th International Conference on Solid-State Sensors, Actuators and Microsystems, TRANSDUCERS 2015).

Research output: Chapter in Book/Report/Conference proceeding › Conference contribution

Guan, W, Chen, L, Rane, T, Kaushik, A & Wang, JTH 2015, Digital droplet ELOHA for nucleic acid molecule counting and analysis. in 2015 Transducers - 2015 18th International Conference on Solid-State Sensors, Actuators and Microsystems, TRANSDUCERS 2015., 7180960, 2015 Transducers - 2015 18th International Conference on Solid-State Sensors, Actuators and Microsystems, TRANSDUCERS 2015, Institute of Electrical and Electronics Engineers Inc., pp. 460-463, 18th International Conference on Solid-State Sensors, Actuators and Microsystems, TRANSDUCERS 2015, Anchorage, United States, 6/21/15. https://doi.org/10.1109/TRANSDUCERS.2015.7180960

Guan W, Chen L, Rane T, Kaushik A, Wang JTH. Digital droplet ELOHA for nucleic acid molecule counting and analysis. In 2015 Transducers - 2015 18th International Conference on Solid-State Sensors, Actuators and Microsystems, TRANSDUCERS 2015. Institute of Electrical and Electronics Engineers Inc. 2015. p. 460-463. 7180960. (2015 Transducers - 2015 18th International Conference on Solid-State Sensors, Actuators and Microsystems, TRANSDUCERS 2015). doi: 10.1109/TRANSDUCERS.2015.7180960

Guan, Weihua ; Chen, Liben ; Rane, Tushar et al. / Digital droplet ELOHA for nucleic acid molecule counting and analysis. 2015 Transducers - 2015 18th International Conference on Solid-State Sensors, Actuators and Microsystems, TRANSDUCERS 2015. Institute of Electrical and Electronics Engineers Inc., 2015. pp. 460-463 (2015 Transducers - 2015 18th International Conference on Solid-State Sensors, Actuators and Microsystems, TRANSDUCERS 2015).

@inproceedings{c7fbcd80038b440d9058ca3afaca588f,

title = "Digital droplet ELOHA for nucleic acid molecule counting and analysis",

abstract = "We present a droplet digital Enzyme-Linked Oligonucleotide Hybridization Assay (droplet digital ELOHA) for reference-free absolute quantification of nucleic acids. Droplet digital ELOHA is able to accurately quantify as few as several hundred copies of nucleic acids in a large, mL-sized sample volume (equivalent to 1 aM or lower) without molecular replication. Droplet digital ELOHA provides a linear response to the number of copies present in a sample to allow for detecting a small fold change unattainable by real time quantitative PCR. Its capability to cope with a large sample volume benefits from the use of magnetic beads to streamline capturing of nucleic acids from the sample and subsequent surface hybridization reactions in droplets. Droplet digital ELOHA is particularly attractive for RNA quantification since it does not require reverse transcription. The absolute quantification ability of droplet digital ELOHA is demonstrated with the analysis of clinical Neisseria Gonorrhoeae 16S rRNA to show its potential diagnostic value.",

keywords = "digital assay, Droplet microfluidics, single molecule counting",

author = "Weihua Guan and Liben Chen and Tushar Rane and Aniruddha Kaushik and Wang, {Jeff Tza Huei}",

note = "Publisher Copyright: {\textcopyright} 2015 IEEE.; 18th International Conference on Solid-State Sensors, Actuators and Microsystems, TRANSDUCERS 2015 ; Conference date: 21-06-2015 Through 25-06-2015",

year = "2015",

month = aug,

day = "5",

doi = "10.1109/TRANSDUCERS.2015.7180960",

language = "English (US)",

series = "2015 Transducers - 2015 18th International Conference on Solid-State Sensors, Actuators and Microsystems, TRANSDUCERS 2015",

publisher = "Institute of Electrical and Electronics Engineers Inc.",

pages = "460--463",

booktitle = "2015 Transducers - 2015 18th International Conference on Solid-State Sensors, Actuators and Microsystems, TRANSDUCERS 2015",

}

TY - GEN

T1 - Digital droplet ELOHA for nucleic acid molecule counting and analysis

AU - Guan, Weihua

AU - Chen, Liben

AU - Rane, Tushar

AU - Kaushik, Aniruddha

AU - Wang, Jeff Tza Huei

PY - 2015/8/5

Y1 - 2015/8/5

N2 - We present a droplet digital Enzyme-Linked Oligonucleotide Hybridization Assay (droplet digital ELOHA) for reference-free absolute quantification of nucleic acids. Droplet digital ELOHA is able to accurately quantify as few as several hundred copies of nucleic acids in a large, mL-sized sample volume (equivalent to 1 aM or lower) without molecular replication. Droplet digital ELOHA provides a linear response to the number of copies present in a sample to allow for detecting a small fold change unattainable by real time quantitative PCR. Its capability to cope with a large sample volume benefits from the use of magnetic beads to streamline capturing of nucleic acids from the sample and subsequent surface hybridization reactions in droplets. Droplet digital ELOHA is particularly attractive for RNA quantification since it does not require reverse transcription. The absolute quantification ability of droplet digital ELOHA is demonstrated with the analysis of clinical Neisseria Gonorrhoeae 16S rRNA to show its potential diagnostic value.

AB - We present a droplet digital Enzyme-Linked Oligonucleotide Hybridization Assay (droplet digital ELOHA) for reference-free absolute quantification of nucleic acids. Droplet digital ELOHA is able to accurately quantify as few as several hundred copies of nucleic acids in a large, mL-sized sample volume (equivalent to 1 aM or lower) without molecular replication. Droplet digital ELOHA provides a linear response to the number of copies present in a sample to allow for detecting a small fold change unattainable by real time quantitative PCR. Its capability to cope with a large sample volume benefits from the use of magnetic beads to streamline capturing of nucleic acids from the sample and subsequent surface hybridization reactions in droplets. Droplet digital ELOHA is particularly attractive for RNA quantification since it does not require reverse transcription. The absolute quantification ability of droplet digital ELOHA is demonstrated with the analysis of clinical Neisseria Gonorrhoeae 16S rRNA to show its potential diagnostic value.

KW - digital assay

KW - Droplet microfluidics

KW - single molecule counting

UR - http://www.scopus.com/inward/record.url?scp=84955518086&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84955518086&partnerID=8YFLogxK

U2 - 10.1109/TRANSDUCERS.2015.7180960

DO - 10.1109/TRANSDUCERS.2015.7180960

M3 - Conference contribution

AN - SCOPUS:84955518086

T3 - 2015 Transducers - 2015 18th International Conference on Solid-State Sensors, Actuators and Microsystems, TRANSDUCERS 2015

SP - 460

EP - 463

BT - 2015 Transducers - 2015 18th International Conference on Solid-State Sensors, Actuators and Microsystems, TRANSDUCERS 2015

PB - Institute of Electrical and Electronics Engineers Inc.

T2 - 18th International Conference on Solid-State Sensors, Actuators and Microsystems, TRANSDUCERS 2015

Y2 - 21 June 2015 through 25 June 2015

ER -

Digital droplet ELOHA for nucleic acid molecule counting and analysis

Abstract

Publication series

Other

Keywords

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

Cite this