DocumentCode :
3462177
Title :
Digital droplet ELOHA for nucleic acid molecule counting and analysis
Author :
Weihua Guan ; Liben Chen ; Rane, Tushar ; Kaushik, Aniruddha ; Wang, Jeff Tza-Huei
Author_Institution :
Dept. of Biomed. Eng., Johns Hopkins Univ., Baltimore, MN, USA
fYear :
2015
fDate :
21-25 June 2015
Firstpage :
460
Lastpage :
463
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 :
RNA; bioMEMS; drops; enzymes; microfluidics; molecular biophysics; RNA quantification; clinical Neisseria Gonorrhoeae 16S rRNA; digital droplet ELOHA; droplet digital enzyme-linked oligonucleotide hybridization assay; magnetic beads; nucleic acid molecule counting; potential diagnostic value; real time quantitative PCR; reference-free absolute quantification; streamline capturing; surface hybridization reactions; Biochemistry; DNA; Dynamic range; Magnetic resonance imaging; Microfluidics; RNA; Testing; Droplet microfluidics; digital assay; single molecule counting;
fLanguage :
English
Publisher :
ieee
Conference_Titel :
Solid-State Sensors, Actuators and Microsystems (TRANSDUCERS), 2015 Transducers - 2015 18th International Conference on
Conference_Location :
Anchorage, AK
Type :
conf
DOI :
10.1109/TRANSDUCERS.2015.7180960
Filename :
7180960
Link To Document :
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