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.