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Amplification of the meningococcal porB gene for non-culture serotype characterization

Published online by Cambridge University Press:  01 June 1998

R. URWIN ,
E. B. KACZMARSKI ,
M. GUIVER ,
A. J. FOX  and
M. C. J. MAIDEN
R. URWIN
Affiliation:
Public Health Laboratory, Withington Hospital, Nell Lane, West Didsbury, Manchester, M20 2LR, UK Present address: Wellcome Trust Centre for the Epidemiology of Infectious Disease, Department of Zoology, University of Oxford, South Parks Road, Oxford, OX1 3PS, UK.
E. B. KACZMARSKI
Affiliation:
Public Health Laboratory, Withington Hospital, Nell Lane, West Didsbury, Manchester, M20 2LR, UK
M. GUIVER
Affiliation:
Public Health Laboratory, Withington Hospital, Nell Lane, West Didsbury, Manchester, M20 2LR, UK
A. J. FOX
Affiliation:
Public Health Laboratory, Withington Hospital, Nell Lane, West Didsbury, Manchester, M20 2LR, UK
M. C. J. MAIDEN
Affiliation:
Wellcome Trust Centre for the Epidemiology of Infectious Disease, Department of Zoology, University of Oxford, South Parks Road, Oxford, OX1 3PS, UK
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Abstract

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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 non-culture 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.

Type
Research Article
Information
Epidemiology & Infection , Volume 120 , Issue 3 , June 1998 , pp. 257 - 262
Copyright
1998 Cambridge University Press