IDEXX: Colisure: Coliform/E. coli Enzyme substrate test; CPRG-MUG test
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Official Method Name
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Colisure Test Kit Procedure |
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Current Revision
| Feb. 28, 1994; current instructions with product |
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Media
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WATER |
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Instrumentation
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Most Probable Number |
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Method Subcategory
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Microbiological |
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Method Source
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Citation
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Brief Method Summary
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Presence/Absence: Add one packet of powder to the 100 ml water sample in a sterile, non-fluorescing vessel. Cap and shake the vessel to dissolve the powder. Incubate at 35C for 24 h. Read the results at 24 h (48 h maximum). If yellow color, the test is negative. If the sample has a red to magenta color, the presence of total coliforms is confirmed. If red to magneta, check for fluorescence by placing a 6W, 365 nm UV light within 5 inches of the sample. If blue fluorescence is observed, the presence of E. coli is confirmed. Quantification: See Standard Methods 20th ed. for MPN analysis (5,10,15 tube serial dilution) or pour the sample reagent mixture directly into a Quanti-Tray and seal according to instructions. Incubate and read the results as indicated above. Count the number of positive red-magenta wells and the number of red-magenta and blue-fluorescent wells. Determine the MPN/100ml using the table provided with the Quanti-Tray. For details, see the package insert. Requirements: Colisure kit from IDEXX. Sterile pipettes (for dilution only). Sterile, unbuffered water for dilutions. Incubator for 35oC controlled within 0.5oC. Long wavelength (365 nm) ultraviolet lamp for fluorescence readings. Cost of analysis: Presence/Absence approx. $4.00; Quantification approx. $5.00. |
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Scope and Application
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Ambient, compliance monitoring (Standard Method 9223B): non-compliance: freshwater (surface & ground), drinking waters EPA Fed Reg (Aug 2001) only for E. coli in ambient waters: fresh surface waters; applicability must be demonstrated for other matrices. Approved for presence/absence of total coliforms and E. coli in drinking water as an alternate test procedure. Approved for presence/absence of total coliforms and E. coli in drinking water as an alternate test procedure. USEPA. 1999 (December 1). National primary and secondary drinking water regulations: analytical methods for chemical and microbiological contaminants and revisions to laboratory certification requirements; final rule. Fed. Reg. 64(230)67449-67467, USEPA (Aug. 30, 2001). Analytical methods for biological pollutants in ambient water; proposed rule. Fed. Reg. 66(169)45811-45829. Clean Water Act section 401. 40 CFR 136.1(c). (state certification, licenses) for compliance monitoring in programs 303(c), 304(a), and 501(a). 136.3 Identification of test procedures. |
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Applicable Concentration Range
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For samples in wells (excluding dilution factors): 1 to 2419 MPN/100 mL for 97-well Quanti-Tray; 1 to 200 MPN/100 mL for 51-well Quanti-Tray |
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Interferences
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Heterotrophs at greater than 2 million/100ml |
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Quality Control Requirements
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Control cultures: Conducted on each lot of Colisure received as a minimum. Inoculate sterile water with either ATCC bacteria or Quanti-Cult (IDEXX)- E. coli (color & fluorescence), K. pneumoniae (color only) and P. aeruginosa (no color or fluorescence). |
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Sample Handling
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Sample preservation: chilled, 1oC to 4oC; 0.0008% (w/w) Na2S2O3 added to chlorinated waters EPA Fed Reg (Aug 2001). 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. Sections 9020B8 and B9, 9060A and B. Sample processing time: less than 5 minutes. |
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Maximum Holding Time
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Sample 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). |
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Relative Cost
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Less than $50 |
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Sample Preparation Methods
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