Chapter title |
Pyrosequencing
|
---|---|
Chapter number | 24 |
Book title |
Pyrosequencing
|
Published in |
Methods in molecular biology, January 2015
|
DOI | 10.1007/978-1-4939-2715-9_24 |
Pubmed ID | |
Book ISBNs |
978-1-4939-2714-2, 978-1-4939-2715-9
|
Authors |
Engström, Anna, Juréen, Pontus, Anna Engström, Pontus Juréen |
Abstract |
Tuberculosis (TB) remains a global health problem. The increasing prevalence of drug-resistant Mycobacterium tuberculosis, the causative agent of TB, demands new measures to combat the situation. Rapid and accurate diagnosis of the pathogen and its drug susceptibility pattern is essential for timely initiation of optimal treatment, and, ultimately, control of the disease. We have developed a molecular method for detection of first- and second-line drug resistance in M. tuberculosis by Pyrosequencing(®). The method consists of seven Pyrosequencing assays for the detection of mutations in the genes or promoter regions, which are most commonly responsible for resistance to the drugs rifampicin, isoniazid, ethambutol, amikacin, kanamycin, capreomycin, and fluoroquinolones. The method was validated on clinical isolates and it was shown that the sensitivity and specificity of the method were comparable to those of Sanger sequencing. In the protocol in this chapter we describe the steps necessary for setting up and performing Pyrosequencing for M. tuberculosis. The first part of the protocol describes the assay development and the second part of the protocol describes utilization of the method. |
X Demographics
Geographical breakdown
Country | Count | As % |
---|---|---|
United Kingdom | 1 | 100% |
Demographic breakdown
Type | Count | As % |
---|---|---|
Members of the public | 1 | 100% |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
Unknown | 12 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Student > Ph. D. Student | 4 | 33% |
Other | 3 | 25% |
Researcher | 1 | 8% |
Student > Master | 1 | 8% |
Unknown | 3 | 25% |
Readers by discipline | Count | As % |
---|---|---|
Agricultural and Biological Sciences | 3 | 25% |
Biochemistry, Genetics and Molecular Biology | 2 | 17% |
Medicine and Dentistry | 2 | 17% |
Business, Management and Accounting | 1 | 8% |
Immunology and Microbiology | 1 | 8% |
Other | 1 | 8% |
Unknown | 2 | 17% |