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Next-generation Sequencing

Next-generation Sequencing

The first human genome sequencing project cost $2.7 billion and was declared complete in 2003, after 15 years of international cooperation and collaboration between several research teams and funding agencies. Today, with the advent of next-generation sequencing technologies, the cost and time of sequencing a human genome have dropped over 100 fold.
Next-Generation Sequencing Methods
Although all next-generation methods use different technologies, they all share a set of standard features.

Sanger Sequencing

Sanger Sequencing

DNA sequencing is a fundamental technique that is routinely used in the biological sciences. This method can be applied to a range of questions at different scales - from the sequencing of a cloned DNA fragment or the study of a mutation in a gene up to whole-genome sequencing. However, despite the widespread use of sequencing today, it was not until 1977 that Fredrick Sanger and his collaborators developed the chain-termination method to decode DNA sequences. It relies on the separation of a...

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Updated: Jun 16, 2026

Multiplex Detection of Bacteria in Complex Clinical and Environmental Samples using Oligonucleotide-coupled Fluorescent Microspheres

Multiplex Detection of Bacteria in Complex Clinical and Environmental Samples using Oligonucleotide-coupled Fluorescent Microspheres

Published on: October 23, 2011

Topology-encoded multiplex diagnostics enabled by strand composition-controlled amplification.

Eun Yeong Lee¹, Dong Geun Lee², Ji-Soo Kwon³

¹Department of Biotechnology, Yonsei University, College of Life Science and Biotechnology, Seoul, 03722, Republic of Korea.

Biosensors & Bioelectronics

|June 13, 2026

Summary

This summary is machine-generated.

This study introduces a novel topology-encoded diagnostic method for identifying Fever of Unknown Origin (FUO) pathogens. It overcomes limitations of current multiplex diagnostics by using structural changes instead of spectral signals for accurate pathogen detection.

Keywords:

Coxiella burnetii DNA nanostructure sensors Fever of unknown origin Orientia tsutsugamushi Primer-blocked asymmetric amplification Topology-encoded diagnostics

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Single Droplet Digital Polymerase Chain Reaction for Comprehensive and Simultaneous Detection of Mutations in Hotspot Regions

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Single Droplet Digital Polymerase Chain Reaction for Comprehensive and Simultaneous Detection of Mutations in Hotspot Regions

Single Droplet Digital Polymerase Chain Reaction for Comprehensive and Simultaneous Detection of Mutations in Hotspot Regions

Published on: September 25, 2018

Area of Science:

Molecular Diagnostics
Nanotechnology
Biotechnology

Background:

Multiplex diagnostics are crucial for identifying Fever of Unknown Origin (FUO) pathogens.
Current platforms face limitations due to spectral overlap and limited optical channels, hindering scalability.

Purpose of the Study:

To develop a topology-encoded diagnostic strategy that shifts multiplexing from spectral to structural space.
To enable target discrimination using electrophoretic mobility signatures, overcoming fluorescence limitations.

Main Methods:

Integration of primer-blocked asymmetric amplification (PBA-amp) with programmable DNA nanostructure sensors (DNA-NS).
Target-specific amplicons induce topological transformations in DNA-NS, generating distinct electrophoretic mobility states.
Multiplex identification via gel-resolved readouts.

Main Results:

Demonstrated duplex detection of Coxiella burnetii and Orientia tsutsugamushi with high sensitivity (9.76 aM) and specificity.
Clinical validation showed 96.4% positive percent agreement and 100% negative percent agreement with real-time PCR.
Discordant cases were limited to low-titer samples near the stochastic limit (Ct > 36).

Conclusions:

Established a structurally encoded framework for nucleic acid diagnostics, decoupling multiplexing from spectral constraints.
Offers a scalable alternative to fluorescence-limited multiplexing with simplified readout.
Provides a promising approach for accurate and efficient pathogen identification in FUO cases.