Amplification of the meningococcal porB gene for non-culture serotype characterization.
Since 1992, the proportion of culture-confirmed meningococcal infections compared with numbers of notified cases of meningococcal disease has decreased in England and Wales. As most meningococcal strain characterization methods depend on a clinical isolate, this has resulted in a loss of epidemiological information. To address this problem, and to aid nonculture diagnosis, a semi-nested PCR protocol for the amplification of the meningococcal porB gene from clinical specimens was developed. This gene encodes the meningococcal serotyping antigen; strain typing data was provided by hybridization of allele-specific oligonucleotide probes to the digoxigenin-labelled porB amplicon in a 'PCR ELISA'. This assay was specific for meningococcal DNA and sensitivities of 0.81 for cerebrospinal fluid (CSF), 0.57 for serum, and 0.62 for whole blood taken from patients with proven meningococcal infection were obtained.
Public Health Laboratory, Withington Hospital, West Didsbury, Manchester, UK.
R
R
Urwin
E B
EB
Kaczmarski
M
M
Guiver
A J
AJ
Fox
M C
MC
Maiden
D001425:Bacterial Outer Membrane Proteins / Q000235:genetics*; D004269:DNA, Bacterial / Q000032:analysis; D004797:Enzyme-Linked Immunosorbent Assay; D009345:Neisseria meningitidis / Q000145:classification* / Q000235:genetics; D015345:Oligonucleotide Probes; D016133:Polymerase Chain Reaction; D018272:Porins; D012703:Serotyping
D001425:Bacterial Outer Membrane Proteins; D004269:DNA, Bacterial; D015345:Oligonucleotide Probes; D018272:Porins; C057699:porin protein, Neisseria