Combined real-time PCR and pyrosequencing strategy for objective, sensitive, specific, and high-throughput identification of reduced susceptibility to penicillins in Neisseria meningitidis.
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.
Thulin
Sara
S
National Reference Laboratory for Pathogenic Neisseria, Department of Clinical Microbiology, Orebro University Hospital, SE-701 85 Orebro, Sweden. sara.thulin@orebroll.se
Olcén
Per
P
Fredlund
Hans
H
Unemo
Magnus
M
D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics; D001483:Base Sequence; D006801:Humans; D008826:Microbial Sensitivity Tests / Q000379:methods; D008969:Molecular Sequence Data; D009345:Neisseria meningitidis / Q000187:drug effects*; D010403:Penicillin Resistance; D046915:Penicillin-Binding Proteins / Q000737:chemistry / Q000235:genetics; D010406:Penicillins / Q000494:pharmacology*; D016133:Polymerase Chain Reaction / Q000379:methods; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA
D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D046915:Penicillin-Binding Proteins; D010406:Penicillins