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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.

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Updated: Jun 13, 2026

Pyrosequencing: A Simple Method for Accurate Genotyping

Pyrosequencing: A Simple Method for Accurate Genotyping

Published on: January 8, 2008

Rapid analysis of resistant mutant genotypes using pyrosequencing.

Catherine Arnold¹

¹Centre for Infections, Health Protection Agency, London, UK.

Methods in Molecular Biology (Clifton, N.J.)

|April 20, 2010

Summary

This summary is machine-generated.

Rapidly detecting drug resistance in Mycobacterium tuberculosis is crucial. New methods combine PCR and Pyrosequencing to quickly identify multidrug-resistant (MDR) strains by analyzing rpoB gene mutations.

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Last Updated: Jun 13, 2026

Pyrosequencing: A Simple Method for Accurate Genotyping

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Published on: January 8, 2008

Pyrosequencing for Microbial Identification and Characterization

Pyrosequencing for Microbial Identification and Characterization

Published on: August 22, 2013

Genotyping Single Nucleotide Polymorphisms in the Mitochondrial Genome by Pyrosequencing

Genotyping Single Nucleotide Polymorphisms in the Mitochondrial Genome by Pyrosequencing

Published on: February 10, 2023

Area of Science:

Microbiology
Genetics
Molecular Biology

Background:

Mycobacterium tuberculosis infects a third of the global population and is a slow-growing organism.
Emergence of multidrug-resistant (MDR) strains of M. tuberculosis necessitates rapid detection for effective treatment and control.
Rifampicin resistance in M. tuberculosis is primarily caused by single nucleotide polymorphisms (SNPs) in the 81-bp rifampicin resistance-determining region (RRDR) of the rpoB gene.

Purpose of the Study:

To develop and validate a rapid, high-throughput method for detecting rifampicin resistance in M. tuberculosis.
To enable timely identification of MDR M. tuberculosis strains for improved patient management and public health interventions.

Main Methods:

Polymerase Chain Reaction (PCR) amplification of the 81-bp rpoB RRDR.
Rapid short-read sequencing using Pyrosequencing technology.
Detection of single nucleotide polymorphisms (SNPs) within the amplified rpoB region.

Main Results:

Successful amplification of the rpoB RRDR from clinical samples (sputum) and early cultures.
High-throughput detection of specific SNPs associated with rifampicin resistance.
Demonstration of the method's ability to rapidly identify MDR M. tuberculosis strains.

Conclusions:

Combining PCR and Pyrosequencing offers a rapid and effective approach for detecting drug resistance-conferring mutations in M. tuberculosis.
This method facilitates high-throughput screening, crucial for controlling the spread of MDR M. tuberculosis.
Early and accurate detection of MDR M. tuberculosis is vital for guiding treatment strategies and preventing further transmission.