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Detection of E. coli in beach water within 1 hour using immunomagnetic separation and ATP bioluminescence
Abstract:The contamination of beach waters occurs from the discharge of storm water and sanitary sewer over?ows containing faecal material. Additional faecal material derives from discharge of animals and waterfowl. In order to protect public from exposure to faecal‐contaminated water, it is required to test enteric indicators in beach water. The problem is that the traditional culture‐based methods cannot meet this goal because it takes too long (>24 h), so the results are not available until a day later. A rapid method for testing beach water for Escherichia coli within 1 h has been developed. Immunomagnetic separation (IMS) and ATP bioluminescence were used for selective capture and quanti?cation, respectively. This rapid method was compared to the current method (m‐TEC) using beach water samples. The beach samples were pre?ltered with a 20 µm pore size ?lter in order to remove algae, plant debris and large particles. The results showed that the pre?ltration step did not trap the bacteria which were present in the original water samples. The pre?ltered water was then passed through a 0.45 µm pore size ?lter for concentration. The deposited bacteria were resuspended and then mixed with superparamagnetic polystyrene beads (diameter of 0.6 µm) that were coated with E. coli antibodies. After IMS, the quanti?cation of the E. coli was done by ATP bioluminescence. The results obtained with IMS‐ATP bioluminescence correlated well with the plate count method (Rsq = 0.93). The detection limit of the assay was about 20 CFU/100 mL, which is well below the US EPA limits for recreational water. The entire procedure can be completed in less than 1 hour. The necessary equipment is portable and was tested on‐site. Copyright © 2004 John Wiley & Sons, Ltd.
Keywords:Escherichia coli  beach  immunomagnetic separation  ATP bioluminescence  rapid method
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