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Journal of Clinical Microbiology, December 2005, p. 6007-6014, Vol. 43, No. 12
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.12.6007-6014.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Multilocus Sequence Typing of Neisseria meningitidis Directly from Clinical Samples and Application of the Method to the Investigation of Meningococcal Disease Case Clusters

Andrew Birtles,1 Katie Hardy,2 Stephen J. Gray,1 Suzanne Handford,1 Edward B. Kaczmarski,1 Valerie Edwards-Jones,3 and Andrew J. Fox1*

Health Protection Agency, Meningococcal Reference Unit, North West Regional Laboratory, Manchester,1 Health Protection Agency, West Midlands Regional Laboratory, Birmingham,2 Department of Biological Sciences, Manchester Metropolitan University, Manchester, United Kingdom3

Received 25 June 2005/ Returned for modification 25 August 2005/ Accepted 27 September 2005

Infections associated with Neisseria meningitidis are a major public health problem in England, Wales, and Northern Ireland. Currently, over 40% of cases are confirmed directly from clinical specimens using PCR-based methodologies without an organism being isolated. A nested/seminested multilocus sequence typing (MLST) system was developed at the Health Protection Agency Meningococcal Reference Unit to allow strain characterization beyond the serogroup for cases confirmed by PCR only. This system was evaluated on a panel of 20 meningococcus-positive clinical specimens (3 cerebrospinal fluid and 17 blood samples) from different patients containing various concentrations of meningococcal DNA that had corresponding N. meningitidis isolates. In each case, the sequence type generated from the clinical specimens matched that produced from the corresponding N. meningitidis isolate; the sensitivity of the MLST system was determined to be less than 12 genome copies per PCR. The MLST system was then applied to 15 PCR meningococcus-positive specimens (2 cerebrospinal fluid and 13 blood samples), each from a different patient, involved in three case clusters (two serogroup B and one serogroup W135) for which no corresponding N. meningitidis organisms had been isolated. In each case, an MLST sequence type was generated, allowing the accurate assignment of individual cases within each of the case clusters. In summary, the adaptation of the N. meningitidis MLST to a sensitive nested/seminested format for strain characterization directly from clinical specimens provides an important tool for surveillance and management of meningococcal infection.

* Corresponding author. Mailing address: Health Protection Agency, Manchester Medical Microbiology Partnership, P.O. Box 209, Manchester Royal Infirmary, Oxford Rd., Manchester M13 9WZ, United Kingdom. Phone: 44 (0)16 1276 5689. Fax: 44 (0)16 1276 5744. E-mail: andrew.fox{at}hpa.org.uk.

Journal of Clinical Microbiology, December 2005, p. 6007-6014, Vol. 43, No. 12
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.12.6007-6014.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

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