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

Schalch, B.; Bader, Lutz; Schau, H.-P.; Bergmann, Rolf; Rometsch, Andrea; Maydl, Gertraud; Kessler, S

doi:10.1128/jcm.41.2.892-895.2003

Cited by 20 publications

(8 citation statements)

References 22 publications

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“…Some Clostridium strains are known to produce extracellular DNase, which may limit the use of DNA fingerprinting methods such as pulsed-field gel electrophoresis (PFGE) (11,25,41,44,45). Since all strains studied were typeable by AFLP, the AFLP method seemed to overcome the problem of extracellular DNase production.…”

Section: Discussionmentioning

confidence: 99%

Identification of Clostridium Species and DNA Fingerprinting of Clostridium perfringens by Amplified Fragment Length Polymorphism Analysis

et al. 2006

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An amplified fragment length polymorphism (AFLP) method was applied to 129 strains representing 24 different Clostridium species, with special emphasis on pathogenic clostridia of medical or veterinary interest, to assess the potential of AFLP for identification of clostridia. In addition, the ability of the same AFLP protocol to type clostridia at the strain level was assessed by focusing on Clostridium perfringens strains. All strains were typeable by AFLP, so the method seemed to overcome the problem of extracellular DNase production. AFLP differentiated all Clostridium species tested, except for Clostridium ramosum and Clostridium limosum, which clustered together with a 45% similarity level. Other Clostridium species were divided into species-specific clusters or occupied separate positions. Wide genetic diversity was observed among Clostridium botulinum strains, which were divided into seven species-specific clusters. The same AFLP protocol was also suitable for typing C. perfringens at the strain level. A total of 29 different AFLP types were identified for 37 strains of C. perfringens; strains initially originating from the same isolate showed identical fingerprinting patterns and were distinguished from unrelated strains. AFLP proved to be a highly reproducible, easy-toperform, and relatively fast method which enables high throughput of samples and can serve in the generation of identification libraries. These results indicate that the AFLP method provides a promising tool for the identification and characterization of Clostridium species.The genus Clostridium is a heterogeneous group of bacteria which currently consists of 181 described species. Clostridia are widely distributed in the environment as well as in the intestinal tract of humans and of many animals. Several Clostridium species are pathogenic to humans, domestic animals, or wildlife and are responsible for well-known clostridial diseases such as tetanus, gas gangrene, botulism, pseudomembranous colitis, and food-borne illness (10). In addition, clostridia can be involved in a variety of human infections, such as cholecystitis, pneumonia, bacteremia, empyema, and abscesses, and can thus be isolated from various clinical specimens. However, many of the isolates can be occasional contaminants, or nonpathogenic clostridia, and may not be involved in the disease process. Therefore, the reliable identification of clostridia, isolated from clinical specimens, is important. In addition, a link must be established between isolated clostridia and pathological changes.Despite the clinical significance of clostridia, reliable, practical, and fast identification methods are few. Although simple tests can serve to identify most commonly isolated Clostridium species, the identification of other clostridia by conventional biochemical testing and gas-liquid chromatography is still laborious, expensive, and time-consuming. Furthermore, several commercial identification systems for anaerobic bacteria have failed to accurately identify Clostridium species (23,...

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Section: Discussionmentioning

confidence: 99%

Identification of Clostridium Species and DNA Fingerprinting of Clostridium perfringens by Amplified Fragment Length Polymorphism Analysis

et al. 2006

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“…In order to solve this problem, we fixed Clostridium cells with formaldehyde. This approach had been previously used to inactivate bacterial DNases and to prevent C. botulinum DNA degradation (Hielm et al, 1998a) and C. perfringens (Schalch et al, 2003) in PFGE protocols. On the other hand, we included thiourea in our PFGE protocol.…”

Section: Modification Of the Pfge Protocolmentioning

confidence: 99%

Enhanced PFGE protocol to study the genomic diversity of Clostridium spp. isolated from Manchego cheeses with late blowing defect

et al. 2012

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“…Reliable whole-cell-to-subtype methodology, obviating the need for DNA purification, would represent a much-VOL. 44,2006 SUBTYPING OF C. PERFRINGENS 1071 needed advancement in the field of DNA-based molecular subtyping.…”

Section: Discussionmentioning

confidence: 99%

“…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.…”

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confidence: 99%

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

et al. 2006

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Clostridium perfringens strains (type A) isolated from an integrated poultry operation were subtyped using repetitive-element PCR with Dt primers. Isolates were obtained from fecal, egg shell, fluff, and carcass rinse samples as part of a previously reported temporally linked epidemiological survey. A total of 48 isolates of C. perfringens were obtained from different stages of the broiler chicken production chain from two separate breeder farms that supplied a single hatchery that in turn provided chicks to a single grow-out farm whose flocks were processed at a single plant. All 48 isolates were typeable (100% typeability) by repetitive-element PCR with Dt primers. This subtyping method was highly reproducible and discriminatory. By repetitive-element PCR with Dt primers, isolates were classified into four major branches with 12 subgroups or clades. The Simpson's index of discrimination was calculated to be 0.96 for groupings of >95% correlation. Toxin gene profiles of the isolates indicated that all of the isolates were C. perfringens alpha-toxin gene positive and 46 of 48 isolates were beta2-toxin gene positive. All strains were negative for beta-and epsilon-toxin genes. Repetitive sequence-based PCR was found to be a technically practical and reproducible means of subtyping C. perfringens libraries from specific epidemiological or production environment settings.

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

Cited by 20 publications

References 22 publications

Identification of Clostridium Species and DNA Fingerprinting of Clostridium perfringens by Amplified Fragment Length Polymorphism Analysis

Identification of Clostridium Species and DNA Fingerprinting of Clostridium perfringens by Amplified Fragment Length Polymorphism Analysis

Enhanced PFGE protocol to study the genomic diversity of Clostridium spp. isolated from Manchego cheeses with late blowing defect

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

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