Detection and genotyping of meningococci using a nested PCR approach
Detection and genotyping of meningococci using a nested PCR approach
An effective vaccine against Neisseria meningitidis serogroup B is required. Outer-membrane protein vaccines have been developed, which may provide protection against common circulating serotypes and serosubtypes in some countries. However, limited genosubtyping data are available because most laboratories use mAbs directed against a limited number of specific serotypes and serosubtypes and laboratories do not genosubtype directly from body fluids due to the lack of a sensitive PCR method. A nested PCR was therefore developed that enables the amplification of the porA gene directly from clinical samples and has the required sensitivity for nucleotide sequencing of the three main variable regions, VR1, VR2 and VR3. Data were compared with those from culture-based nucleotide sequencing, and the use of this method increased the availability of genosubtype information by 45 %, thereby indicating the impact that this methodology has on the data provided and the implications for vaccine design.
Genes, Bacterial/genetics, Genotype, Humans, Meningococcal Infections/blood, Neisseria meningitidis/classification, Phenotype, Polymerase Chain Reaction/methods, Porins/analysis
51-57
Diggle, M A
d739c25b-e038-4a9a-850d-2e2fd6ff4a4f
Clarke, S C
f7d7f7a2-4b1f-4b36-883a-0f967e73fb17
January 2003
Diggle, M A
d739c25b-e038-4a9a-850d-2e2fd6ff4a4f
Clarke, S C
f7d7f7a2-4b1f-4b36-883a-0f967e73fb17
Diggle, M A and Clarke, S C
(2003)
Detection and genotyping of meningococci using a nested PCR approach.
Journal of Medical Microbiology, 52 (Pt 1), .
(doi:10.1099/jmm.0.05032-0).
Abstract
An effective vaccine against Neisseria meningitidis serogroup B is required. Outer-membrane protein vaccines have been developed, which may provide protection against common circulating serotypes and serosubtypes in some countries. However, limited genosubtyping data are available because most laboratories use mAbs directed against a limited number of specific serotypes and serosubtypes and laboratories do not genosubtype directly from body fluids due to the lack of a sensitive PCR method. A nested PCR was therefore developed that enables the amplification of the porA gene directly from clinical samples and has the required sensitivity for nucleotide sequencing of the three main variable regions, VR1, VR2 and VR3. Data were compared with those from culture-based nucleotide sequencing, and the use of this method increased the availability of genosubtype information by 45 %, thereby indicating the impact that this methodology has on the data provided and the implications for vaccine design.
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Published date: January 2003
Keywords:
Genes, Bacterial/genetics, Genotype, Humans, Meningococcal Infections/blood, Neisseria meningitidis/classification, Phenotype, Polymerase Chain Reaction/methods, Porins/analysis
Identifiers
Local EPrints ID: 455316
URI: http://eprints.soton.ac.uk/id/eprint/455316
ISSN: 0022-2615
PURE UUID: 91ca974a-00a3-4f78-a2be-5ca33c2a6245
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Date deposited: 16 Mar 2022 18:07
Last modified: 17 Mar 2024 03:07
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Author:
M A Diggle
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