Colilert Test Kit Procedure

Introduction and Product Use 5. Aseptically cap and seal the vessel.
Colilert Reagent is used for the simultaneous detection and confirmation 6. Shake until dissolved.
of total coliforms and E.coli in water. It is based on IDEXX's patented 7. Incubate for 24 hours at 35° ± 0,5°C.
Defined Substrate Technology® (DST™) This product utilizes nutrient
8. Read the results at 24 hours. Compare each result against the
indicators that produce color and/or fluorescence when metabolized by comparator dispensed into an identical vessel.
total coliforms and E. coli. When the reagent is added to the sample and - If no yellow color is observed, the test is negative.
incubated, it can detect these bacteria at 1CFU/100 ml within 24 hours
- If the sample has a yellow color equal to or greater than the
with as many as 2 million heterotrophic bacteria/100ml present. This test
comparator, the presence of total coliforms is confirmed. If color is
is not intended for marine water. For marine water use Colilert-18, (IDEXX
not uniform, mix by inversion then recheck.
Catalog #WP020-18 or WP200-18).
- If the sample is yellow, but lighter than the comparator, it may be
Materials incubated an additional 4 hours (but no more than 28 hours total). If
the sample is coliform positive, the color will intensify. If it does not
Catalog # Mfg.-Code Contents
intensify, the sample is negative.
WP020 98-20704-00 20 Snap Packs of reagent for 100 ml - If yellow is observed, check vessel for fluorescence by placing a 6
water samples watt 365 nm UV light within five inches of the sample in a dark
WP200 98-20705-00 200 Snap Packs of reagent for 100 ml environment. Be sure the light is facing away from your eyes and
water samples towards the vessel. If fluorescence is greater or equal to the
W050 98-20708-00 20 Snap Packs of reagent for 50 ml fluorescence of the comparator, the presence of E. coli is confirmed.
water samples
Quanti-Tray™ (MPN) Enumeration Test
W050B 98-20709-00 200 Snap Packs of reagent for 50 ml
water samples Procedure for 100 ml sample
Colilert can be also used for multiple tube MPN analysis (eg: 5-tube, 10-
Storage tube, or 15-tube serial dilutions). Consult US Standard Methods for
Store at 4°- 30°C away from light. appropriate MPN tables. For increased accuracy and counting range, use
the lDEXX Quanti-Tray (catalog # WQT-100) and follow the above
Presence/Absence Test Procedure presence/absence steps one through six and then proceed as follows:
1. Carefully separate one Snap Pack from the 1. Pour the sample reagent mixture from step 6
strip taking care not to accidentally open adjacent above directly into the tray avoiding contact
pack. with the foil tab, and then seal the tray
2. Tap the Snap Pack to ensure that all of the Colilert according to Quanti-Tray instructions.
powder is in the bottom part of the pack.
2. Incubate for 24 hours at 35°±0.5°C.
3. Open one pack by snapping back the top at the
3. Follow the same interpretation directions from step 8 above to count
scoreline as shown.
the number of positive wells. Refer to the MPN table provided with the
Caution: Do not touch the opening of pack. Quanti-Tray to determine the Most Probable Number (MPN) of total
4. Add the reagent to the water sample in a sterile, coliforms (yellow wells) and E. coli (fluorescent wells) in your sample.
transparent, non-fluorescent vessel (100 ml for The color and fluorescence intensity of positive wells may vary.
WP020 and WP200, 50 ml for W050 and W050B).
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 Procedural Notes
1. If an inoculated Colilert sample is inadvertently incubated over 28
hours, the following guidelines apply: Lack of yellow color is a VALID
NEGATIVE TEST. A yellow color after 28 hours is not valid and should
be repeated or verified.
2. Some water samples containing humic material may have an innate
color. If a water sample has some background color, compare
inoculated Colilert sample to a control blank of the same water
sample.
3. Use sterile water, not buffered water for making dilutions. Colilert is
already buffered. Always add Colilert to the proper volume of diluted
sample after making dilutions.
4. Colilert is a primary water test. Colilert performance characteristics do
not apply to samples altered by any pre-enrichment or concentration.

Quality Control Procedure

Quality control should be conducted on each lot of Colilert, or more often
as regulations require. Inoculate sterile water (100ml for WP020 and
WP200, 50 ml for W050 and W050B) with the Quanti-Cult™ or American
Type Culture Collection (ATCC) bacteria listed below. Follow the above
test procedure, and compare test results to the expected results below.
Quanti-Cult Organism ATCC # Expected Result
E. coli 25922 or11775 yellow, fluorescent
Klebsiella pneumoniae 31488 yellow, no fluorescent
Pseudomonas aeruginosa 10145 or 27853 clear, no fluorescent

For IDEXX technical assistance in the US,

call 1-800-321-0207 or 207-856-0300.
For IDEXX technical assistance in the UK,
call 01753-891660.

US Patent No. 4925789. Other patents pending.

Quanti-Cult™ cultures are available from IDEXX - catalog number WKIT-1001.
Quanti-Cult is a trademark of Chrisope Technologies.
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