Immunomagnetic separation of Salmonella from foods and their detection using immunomagnetic particle (IMP)-ELISA.

An immunomagnetic particle based ELISA (IMP-ELISA) for the detection of Salmonella from foods has been developed using Dynabeads anti-Salmonella (Dynal, Oslo, Norway). Appropriate sample preparation protocols to allow rapid detection of Salmonella serovariants in processed (powdered egg products) and non-processed (raw chicken) samples have been established. Pre-enriched broths of heat processed samples likely to harbour only low levels of competitive enteric flora, were boiled and used directly for IMP-ELISA. For non-heat processed or raw samples likely to contain higher numbers of such competing organisms, live Salmonella cells were first isolated by immunomagnetic separation (IMS) from standard pre-enrichment broths, and then post-selectively enriched for a short time in M-broth followed by boiling before IMP-ELISA. The total assay time including sample preparation was under 26 h for both types of procedure, with a lower detection limit of 10(5) Salmonella cells/ml of sample. In an evaluation of naturally contaminated poultry samples, all 45 of 48 samples previously shown to contain salmonellae in a comparison of ISO, IMS-Plating, Salmonella-Tek ELISA (Organon Teknika, Inc. Durham, NC) and a modification of the latter based on IMS, were identified as positive. None of the other methods gave positives for all 45.

Evaluation of methods for the isolation and detection of Escherichia coli O157 in minced beef.

This study has evaluated enrichment and detection procedures for the isolation and detection of Escherichia coli O157 inoculated into minced beef. The use of a 24 h enrichment in modified EC broth containing novobiocin allowed low numbers of contaminating cells to multiply to levels detectable on culture media and by ELISA test kits. Total analysis time was reduced by the use of the Dynabead immunomagnetic separation system. The use of the Petrifilm Test Kit-HEC for E. coli O157:H7 and Organon Teknika EHEC-TEK system detected low numbers of contaminating cells following enrichment and reduced analysis time by 1 d. The incorporation of cefixime and tellurite into Sorbitol MacConkey Agar increased the rate and ease of isolation of E. coli O157 and its use is therefore recommended.

The isolation and detection of Escherichia coli O157 by use of immunomagnetic separation and immunoassay procedures.

The use of immunomagnetic separation (IMS) techniques has been reported to reduce the total test time, and improve the sensitivity, of microbiological tests done on foods. This approach is being adopted in epidemiological investigations into suspected foodborne outbreaks of Escherichia coli O157 infection and has gained acceptance by public health laboratories and the food industry. This study demonstrated the ability of a commercially available IMS procedure, Dynabeads anti-E. coli O157, to enable detection of a few cells of E. coli O157 in 25 g of inoculated minced beef, giving results 1 d earlier than a cultural analysis of similar sensitivity. With correct choice of enrichment broths, IMS may increase isolation rate of E. coli O157 compared to that obtained using conventional cultural methods. It is suggested that this may be due to an increase in relative concentration of E. coli O157 compared with the background microflora present in minced beef, which may reduce reliability of non-IMS detection procedures by masking or mimicking target cells on selective/differential solid media. The use of an immunoassay incorporating an IMS step, EHEC-Tek (Organon-Teknika), enabled detection of a few cells of E. coli O157 in 25 g of minced beef. Comparison of the IMS-ELISA with a standard ELISA procedure (Tecra) indicated the sensitivity of the latter system to be greater, perhaps resulting in the higher isolation rate. The use of a method to reliability isolate and detect extremely low levels of E. coli O157 in a food is necessary to aid reduction in the incidence of this most serious of foodborne pathogens.