Digital droplet ELOHA for nucleic acid molecule counting and analysis

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

    Research output: Chapter in Book/Report/Conference proceedingConference 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 languageEnglish (US)
    Title of host publication2015 Transducers - 2015 18th International Conference on Solid-State Sensors, Actuators and Microsystems, TRANSDUCERS 2015
    PublisherInstitute of Electrical and Electronics Engineers Inc.
    Pages460-463
    Number of pages4
    ISBN (Electronic)9781479989553
    DOIs
    StatePublished - Aug 5 2015
    Event18th International Conference on Solid-State Sensors, Actuators and Microsystems, TRANSDUCERS 2015 - Anchorage, United States
    Duration: Jun 21 2015Jun 25 2015

    Other

    Other18th International Conference on Solid-State Sensors, Actuators and Microsystems, TRANSDUCERS 2015
    CountryUnited States
    CityAnchorage
    Period6/21/156/25/15

    Fingerprint

    Nucleic acids
    nucleic acids
    counting
    Molecules
    oligonucleotides
    enzymes
    molecules
    Oligonucleotides
    Assays
    Enzymes
    beads
    surface reactions
    Surface reactions
    Transcription
    RNA

    All Science Journal Classification (ASJC) codes

    • Instrumentation
    • Electrical and Electronic Engineering

    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). [7180960] Institute of Electrical and Electronics Engineers Inc.. https://doi.org/10.1109/TRANSDUCERS.2015.7180960
    Guan, Weihua ; Chen, Liben ; Rane, Tushar ; Kaushik, Aniruddha ; Wang, Jeff Tza Huei. / 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
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    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.",
    author = "Weihua Guan and Liben Chen and Tushar Rane and Aniruddha Kaushik and Wang, {Jeff Tza Huei}",
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    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, 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

    Digital droplet ELOHA for nucleic acid molecule counting and analysis. / Guan, Weihua; Chen, Liben; Rane, Tushar; Kaushik, Aniruddha; Wang, Jeff Tza Huei.

    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.

    Research output: Chapter in Book/Report/Conference proceedingConference contribution

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    AU - Rane, Tushar

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    AU - Wang, Jeff Tza Huei

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    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.

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    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 https://doi.org/10.1109/TRANSDUCERS.2015.7180960