Immunochemical fingerprinting of Clostridium difficile strains isolated from an outbreak of antibiotic-associated colitis and diarrhoea

Abstract

SUMMARY. Twenty eight strains of Clostridium dificile, isolated from an outbreak of antibiotic-associated colitis and diarrhoea in an orthopaedic ward and from sporadic cases throughout Sweden, were sent to Edinburgh for immunochemical fingerprinting without information about their origin. EDTA extracts of the organisms were examined by crossed immunoelectrophoresis (CIE), polyacrylamide gel electrophoresis (PAGE) and electroblot transfer. Two patterns were revealed by CIE: group A (18 strains) and group B (10 strains). PAGE and electroblot transfer revealed one major group of 10 strains (group l), six small groups of two or three strains and six strains which were unlike any other strain. The CIE group B and PAGE-electroblot group 1 were identical. Nine of the 10 strains in this group were from patients in the outbreak. These findings indicate that a single strain spread in the orthopaedic ward as a nosocomial infection and that this strain differed from most other strains investigated. The PAGE-electroblot technique should, therefore, greatly aid investigations into the epidemiology of C. dificile infections.

Genetic Diversity Among Toxigenic Clostridia Isolated From Soil, Water, Meat and Associated Polluted Sites in South India

“…To type our strains, a simple and straightforward method, originally described by Poxton et al, [12] i.e., the electrophoretic patterns of whole-cell and excretory proteins from strains grown under standardized conditions, was used. By this method, a great variety of patterns was obtained from the strains examined.…”

Identification of outbreak-associated and other strains ofClostridium difficileby numerical analysis of SDS-PAGE protein patterns

“…The immediate environment of infected patients has been found by a number of workers [41,42] to be contaminated with C. difficile, and in some studies, typing has confirmed that environmental isolates are often of the same type as those of the infected patient with which they are associated [14,28]. The Manchester environmental samples included two samples from bed linen and the floor associated with one patient, plus a cubicle curtain sample from a second patient; all were of the same EP type as the patient isolates themselves, i.e.…”

“…Differing electrophoretic methodologies have been employed in the typing of C. difficile by protein patterns and it appears to be the most popular general approach. These include radio-labelled [12,16], EDTA-extracted surface proteins [7,22], and whole-cell proteins [23][24][25] and, in addition, these may be coupled with immunoblotting techniques [26][27][28]. Discrimination is extremely high and in theory all strains can be typed; in practice, well over 95 % of isolates can be typed to previously defined types and any new patterns can be added as new types.…”

“…For epidemiological studies, a variety of methods have been applied to the typing of C. difficile: bacteriophage and bacteriocin typing [11,17,18], serotyping [19][20][21], protein typing (including radio-labelled [12,16], EDTA-extracted surface [7,22], and whole-cell proteins [23][24][25]), immunoblotting techniques [26][27][28], plasmid profiling [29], restriction endonuclease fingerprinting [30,31], restriction fragment length polymorphisms [32], the polymerase chain reaction [33], and pyrolysis mass spectrometry [34]. [35], infection contributed to 17 deaths amongst the mainly geriatric patients.…”

Clostridium difficile in a geriatric unit: a prospective epidemiological study employing a novel S-layer typing method

“…The main aims of this investigation were to determine the level of C. difficile colonization and disease in a population of elderly patients by routine culture and toxin detection methods, and to use a novel typing method based on the variation of the S-layer proteins from C. difficile to compare the strains in the patients and the ward environment. S-typing is a simplified modification of the method based on EDTA-soluble peptides, which was the first-ever method used to show that C. difficile was acquired exogenously (Poxton et al, 1984). The present study was part of a major prospective epidemiological study to investigate the use of inferential stochastic modelling to determine control methods for C. difficile disease -this is being submitted elsewhere.…”