Genotypic characterization of Neisseria meningitidis using Pyrosequencing
Genotypic characterization of Neisseria meningitidis using Pyrosequencing
Pyrosequencing involves the synthesis of single-stranded deoxyribonucleic acid leading to rapid and accurate analysis of nucleotide sequences. This article describes the development of typing assays for the characterization of Neisseria meningitidis using Pyrosequencing. This involved developing methods for the nucleotide sequence analysis of important variable regions contained on a major capsule gene and on outer membrane protein genes that are used for grouping, typing, and subtyping meningococci. To achieve this, primers were designed for amplification of three genes, siaD, porB, and porA from the four main serogroups B, C, Y, and W135. To facilitate throughput and reproducibility, the method was also automated. Data from 717 isolates have shown that Pyrosequencing can be used for the single nucleotide polymorphism and sequence-analysis characterization of meningococci.
Bacterial Typing Techniques, DNA, Single-Stranded/chemistry, Genotype, Neisseria meningitidis/genetics, Porins/genetics, Sequence Analysis, DNA/methods
139-45
Diggle, Matthew A
94088f60-f398-42a0-a221-f71978433987
Clarke, Stuart C
f7d7f7a2-4b1f-4b36-883a-0f967e73fb17
October 2004
Diggle, Matthew A
94088f60-f398-42a0-a221-f71978433987
Clarke, Stuart C
f7d7f7a2-4b1f-4b36-883a-0f967e73fb17
Diggle, Matthew A and Clarke, Stuart C
(2004)
Genotypic characterization of Neisseria meningitidis using Pyrosequencing.
Molecular Biotechnology, 28 (2), .
(doi:10.1385/mb:28:2:139).
Abstract
Pyrosequencing involves the synthesis of single-stranded deoxyribonucleic acid leading to rapid and accurate analysis of nucleotide sequences. This article describes the development of typing assays for the characterization of Neisseria meningitidis using Pyrosequencing. This involved developing methods for the nucleotide sequence analysis of important variable regions contained on a major capsule gene and on outer membrane protein genes that are used for grouping, typing, and subtyping meningococci. To achieve this, primers were designed for amplification of three genes, siaD, porB, and porA from the four main serogroups B, C, Y, and W135. To facilitate throughput and reproducibility, the method was also automated. Data from 717 isolates have shown that Pyrosequencing can be used for the single nucleotide polymorphism and sequence-analysis characterization of meningococci.
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Published date: October 2004
Keywords:
Bacterial Typing Techniques, DNA, Single-Stranded/chemistry, Genotype, Neisseria meningitidis/genetics, Porins/genetics, Sequence Analysis, DNA/methods
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Local EPrints ID: 454177
URI: http://eprints.soton.ac.uk/id/eprint/454177
ISSN: 1073-6085
PURE UUID: 47fe3b82-2d60-4f35-9eba-19c231f6cebf
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Date deposited: 01 Feb 2022 17:57
Last modified: 17 Mar 2024 03:07
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Author:
Matthew A Diggle
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