Combined real-time PCR and pyrosequencing strategy for objective, sensitive, specific, and high-throughput identification of reduced susceptibility to penicillins in Neisseria meningitidis

Sara Thulin; Per Olcén; Hans Fredlund; Magnus Unemo

doi:10.1128/AAC.00914-07

Combined real-time PCR and pyrosequencing strategy for objective, sensitive, specific, and high-throughput identification of reduced susceptibility to penicillins in Neisseria meningitidis

Antimicrob Agents Chemother. 2008 Feb;52(2):753-6. doi: 10.1128/AAC.00914-07. Epub 2007 Dec 10.

Authors

Sara Thulin¹, Per Olcén, Hans Fredlund, Magnus Unemo

Affiliation

¹ National Reference Laboratory for Pathogenic Neisseria, Department of Clinical Microbiology, Orebro University Hospital, SE-701 85 Orebro, Sweden. [email protected]

Abstract

A segment of penA in Neisseria meningitidis strains (n = 127), including two nucleotide sites closely associated to reduced susceptibility to penicillins, was amplified and pyrosequenced. All results were in concordance with Sanger sequencing, and a high correlation between alterations in the two Pen(i)-specific sites and reduced susceptibility to penicillins was identified.

Publication types

Evaluation Study
Research Support, Non-U.S. Gov't

MeSH terms

Anti-Bacterial Agents / pharmacology
Bacterial Proteins / chemistry
Bacterial Proteins / genetics
Base Sequence
Humans
Microbial Sensitivity Tests / methods
Molecular Sequence Data
Neisseria meningitidis / drug effects*
Penicillin Resistance*
Penicillin-Binding Proteins / chemistry
Penicillin-Binding Proteins / genetics
Penicillins / pharmacology*
Polymerase Chain Reaction / methods
Sensitivity and Specificity
Sequence Analysis, DNA

Substances

Anti-Bacterial Agents
Bacterial Proteins
Penicillin-Binding Proteins
Penicillins