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Method Summary

Method Source Micrology Laboratories L.L.C.
Method NumberColiscan
Revision Information1999
Method NameDescriptive Name: E. coli and other coliform bacteria by Coliscan MF

Official Name: Coliscan MF

MediaMedia: WATER
SubcategoryMicrobiological
Citation Tests on improved, simplified membrane filtration method for Escherichia coli detection and enumeration.
Method DownloadThis is a proprietary method.
Please visit the method owner's site for method documentation
Summary of Method100 mL water sample is filtered through a 0.45 micron membrane filter. Medium (1.75 - 2 ml) is dispensed onto a sterile pad in a 50 mm petri pad, or plates may be prepared by dispensing 5 mL of agar-based medium per 50 mm plate. The filter is then transferred to the plate, covered and incubated at 35C for 24 h. The presence of pink colonies is indicative of coliforms; the presence of blue/purple colonies is indicative of E. coli. Requirements: a low power (10-15 magnification) binocular wide-field dissecting microscope or equivalent may be necessary to to provide optimal viewing of the colonies.
Scope and Applicationambient monitoring: all water - surface water, ground water, waste effluents, swimming pool, drinking water
Applicable Concentration Range20 - 80 CFU/100 mL is considered ideal for enumeration. Maximum: 200 CFU/100 mL; dilution is required for samples that exceed this level.
InterferencesWaters containing sediment or suspended materials such as iron and manganese can clog the filter pores and prevent filtration, or mask target colonies and prevent accurate counting. Aeromonas may yield a positive coliform reaction under certain conditions, such as if one of the inhibitors in the medium is eliminated through adverse storage or outdating. Aeromonas does not interfere with E. coli determination. Sources of interference in MF methods (USEPA Fed Reg Aug. 2001): high turbidity, toxic compounds, or large numbers of non-coliform (background) bacteria, and organisms damaged by chlorine or toxic compounds.
QC Requirements(Standard Methods, 20th ed., Section 9020 B8 and B9). Testing should be conducted on every lot of reagent with a known positive and negative strain. Verify filters are free of coliform bacteria.
Sample HandlingChill samples, 1oC to 4oC. Techniques for collection: Standard Methods for the Examination of Water and Wastewater, 20th Edition. L. Clesceri, A. Greenberg, and A. Eaton (editors). APHA: Washington, DC. 1998. Section 9060A
Sample processing time: greater than 10 minutes. To minimize interferences causing underestimation of organism density in MF methods (USEPA Fed Reg Aug. 2001): replicates of smaller sample dilutions/volumes may be filtered and the results combined.
Max Holding TimeSample should be analyzed within 6 h after sampling and within 2 h from receipt of sample in lab for compliance or 24 h for routine monitoring (Standard Methods, 20th ed., Section 9060B); however, a 6 h holding time for all samples is highly recommended (Myers and Sylvester, 1997). Freeze kits for
Relative Cost/EffortLess than $50
Sample Prep Method(s) 
 

Method E. coli and other coliform bacteria by Coliscan MF has 2 analytes associated with it.

Analyte


Detection
Level


Bias Precision Pct False
Positive
E. coli
(68583-22-2)
1 CFU/100 mLN/AN/A 
Total coliforms
(E761700)
1 CFU/100 mLN/AN/A 
export_analytes
Precision Descriptor NotesAeromonas can yield false positives for coliform test under certain conditions, such as if one of the inhibitors in the medium is eliminated through adverse storage or outdating. Aeromonas does not interfere with E. coli determination. General comments (USEPA Fed Reg Aug. 2001): Membrane filter (MF) methods are generally more precise in enumerating target organisms than MPN tests. Based on susceptibility to interferences, MF tests may underestimate the number of viable bacteria.
Detection Level NoteDetermined by using the largest volume plated, and the lowest number of organisms that can be counted. For example, if 100 mL was the largest volume plated, the detection would be 1 colony per 100 mL.
 
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