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Journal of Clinical Microbiology, January 2002, p. 101-104, Vol. 40, No. 1
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.1.101-104.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Molecular Fingerprinting of Clostridium difficile Isolates: Pulsed-Field Gel Electrophoresis versus Amplified Fragment Length Polymorphism

Corné H. W. Klaassen,* Hanneke A. van Haren, and Alfons M. Horrevorts

Department of Medical Microbiology and Regional Public Health Laboratory, Canisius Wilhelmina Hospital, Nijmegen, The Netherlands

Received 25 May 2001/ Returned for modification 7 August 2001/ Accepted 15 October 2001

Two molecular fingerprinting techniques, pulsed-field gel electrophoresis (PFGE) and amplified fragment length polymorphism (AFLP), were used to investigate the epidemiological relatedness among Clostridium difficile isolates from suspected outbreaks in three general hospitals. Analysis by PFGE yielded inconclusive data as a result of extensive DNA degradation. Although this degradation could be prevented to a certain extent by the inclusion of thiourea in the electrophoresis buffer, the weak DNA banding patterns obtained in this way were still far from optimal. AFLP data were obtained by using fluorescently labeled PCR primers and analysis on an ABI PRISM automated DNA analysis platform. AFLP analysis yielded high resolution and highly reproducible DNA fingerprinting patterns from which the epidemiological relatedness among the isolates could easily be determined. AFLP results could be readily obtained within 24 h, whereas 3 to 4 days were routinely required to complete the lengthy PFGE protocol. AFLP clearly proved to be a much more fail-safe fingerprinting method for C. difficile isolates, especially for those isolates for which a standard PFGE procedure yielded inconclusive results due to DNA degradation.

* Corresponding author. Mailing address: Department of Medical Microbiology and Regional Public Health Laboratory, Canisius Wilhelmina Hospital, Weg door Jonkerbos 100, NL-6532 SZ Nijmegen, The Netherlands. Phone: 31-24-3657514. Fax: 31-24-3657516. E-mail: c.klaassen{at}cwz.nl.

Journal of Clinical Microbiology, January 2002, p. 101-104, Vol. 40, No. 1
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.1.101-104.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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