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Amplified fragment length polymorphism (AFLP) analysis of Clostridium novyi, C. perfringens and Bacillus cereus isolated from injecting drug users during 2000

J. MCLAUCHLIN, J.E. SALMON, S. AHMED, J.S. BRAZIER, M.M. BRETT, R.C. GEORGE^* and J. HOOD^||

Food Safety Microbiology Laboratory, Division of Gastrointestinal Infections and *Respiratory and Systemic Infection Laboratory, PHLS Central Public Health Laboratory, 61 Colindale Avenue, London NW9 5HT, Department of Medical Microbiology and Public Health Laboratory and PHLS Anaerobe Reference Unit, University Hospital of Wales, Heath Park, Cardiff CF14 4XW, Greater Glasgow Health Board, 350 Vincent Street, Glasgow G3 8YU and ||Department of Clinical Microbiology, Glasgow Royal Infirmary, 84 Castle Street, Glasgow G4 OSF

Corresponding author: Dr J. McLauchlin (e-mail: jmclauchlin{at}phls.org.uk).

Received 15 May 2002; accepted 20 July 2002.

Abstract

As part of the follow-up investigations associated with an outbreak of severe illness and death among illegal injecting drug users during 2000, 43 cultures of Clostridium novyi type A, 40 C. perfringens type A and 6 isolates of Bacillus cereus were characterised by amplified fragment length polymorphism (AFLP) analysis. Among the 43 C. novyi isolates, 23 different AFLP profiles were detected. The same AFLP profile was detected in isolates from 18 drug users investigated during 2000 from Scotland, England, the Republic of Ireland and Norway and a wound from a patient in 2000 who was not identified as a drug user. Unique AFLP profiles were obtained from four drug users from England and the Republic of Ireland, 10 historical isolates from culture collections, an isolate from food (1989) and three isolates from wounds (1995, 1991, 1988). The 40 C. perfringens isolates were from 13 drug users, the contents of one syringe and two samples of heroin. Sixteen AFLP types of C. perfringens were distinguished and there was little evidence for commonality among the isolates. The AFLP types of C. perfringens from heroin differed and were unique. Six isolates of B. cereus were from four drug users and two samples of heroin. Four different AFLP patterns were distinguished. Three AFLP types were isolated from four drug users. B. cereus isolates from an aspirate and a heroin sample collected from the same drug user were identical, and were also indistinguishable from an isolate from a groin infection in a second drug user. The AFLP type of the isolate from a second and unrelated heroin sample was unique. The AFLP results showed no or very limited evidence for commonality between the different isolates of B. cereus and C. perfringens. In marked contrast, the C. novyi isolates from the majority of the drug users during 2000 were homogeneous, suggesting a common source or clonal selection of a C. novyi type, or both, which either had an adaptive advantage in spore germination, survival or growth following the drug preparation and the injection procedure, or produced a more severe clinical presentation.

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