Molecular methods for the analysis of Clostridium perfringens relevant to food hygiene

Abstract

Clostridium perfringens continues to be a common cause of food-borne disease [1,2]. It produces an enterotoxin (CPE) which is released upon lysis of the vegetative cell during sporulation in the intestinal tract. Catering premises with insufficient cooling and reheating devices often seem to be the cause of outbreaks of C. perfringens food poisoning. Typing of C. perfringens is of great importance for investigating sources of food poisoning cases and for studying the epidemiology of this microorganism. This report describes the examination of 155 C. perfringens isolates by molecular methods. Isolates were taken from 10 food poisoning outbreaks and cases (n = 34, food and fecal isolates) and from meat and fish pastes (n = 121). Isolates were characterized by plasmid profiling, ribotyping, and/or macrorestriction analysis by pulsed-field gel electrophoresis (PFGE). Results show that all three methods are suitable for classifying C. perfringens isolates below the species level. Ribopatterns and PFGE patterns can be interpreted more easily than plasmid profiling results and can be recommended for contamination studies and epidemiologic investigation of food poisonings associated with C. perfringens. ß

Molecular Subtyping of Poultry-Associated Type AClostridium perfringensIsolates by Repetitive-Element PCR

“…Pulsed-field gel electrophoresis (PFGE) has been used for subtyping of Clostridium spp. including C. perfringens (10,13,17,18,22,29,31,37,(43)(44)(45)(46); however, less-thancomplete typeability has been reported. For example, Maslanka et al (31) utilized PFGE (SmaI digestion) for subtyping of C. perfringens food-borne disease outbreak strains and found that of 62 C. perfringens isolates tested, 8% were untypeable by PFGE.…”

Molecular Typing of Clostridium perfringens from a Food-Borne Disease Outbreak in a Nursing Home: Ribotyping versus Pulsed-Field Gel Electrophoresis

“…As C. perfringens is also part of the intestinal flora, molecular typing is important for investigating clonal relationships in outbreaks and for studying the molecular epidemiology of this microorganism. Several molecular methods have been used successfully for C. perfringens strain differentiation; among them are serotyping (12), plasmid isolation (4, 11), sodium dodecyl sulfate-polyacrylamide gel electrophoresis (9a), pulsed-field gel electrophoresis (PFGE) (16,19), and ribotyping (1). We describe the examination of 17 food-borne disease-related C. perfringens isolates by ribotyping (7) and PFGE with three different cell lysis methods.…”

“…We describe the examination of 17 food-borne disease-related C. perfringens isolates by ribotyping (7) and PFGE with three different cell lysis methods. PFGE is presumed to offer greater discriminatory power, typeability, and reproducibility than many other DNA-based typing methods, including ribotyping, and has been applied successfully for genotyping of various C. perfringens isolates (16,19,20). Nevertheless, DNA degradation problems with certain strains due to endogenous bacterial nucleases, which are rather common among clostridial isolates, have been reported (5,7,19,20).…”

“…This shows, in accordance with many researchers who have reported problems with Clostridium sp. DNA degradation (1,8,9,10,16,19), that modified PFGE protocols with improved protection against endonucleases activity need to be developed. However, few results concerning the PFGE typeability of C. perfringens strains involved in food-borne diseases have been published (16; Klein et al, Proc.…”

Potential Role of Anaerobic Bacteria as Fish Pathogens

“…This may increase the health risk to the consumer, as C. perfringens may cause outbreaks of food poisoning associated with the consumption of fish and its products [7]. It produces an enterotoxin which is released upon lysis of the vegetative cell during sporulation in the intestinal tract [8]. C. perfringens causes clinical signs of gastroenteritis, which often resolve within 24 hours with minimal or no treatment.…”