THE COLILERT SYSTEM.
Introduction
This system is used to detects for both total coliforms and E. coli and the results are obtained in
24 hours.
The Colilert system is usually available in:
I. 100ml format
II. 250ml format
III. 10ml formats
The 100ml format
Used to test for:
1) Presence- absence detection of total coliforms and E. coli
2) For quantification of total coliforms and E. coli.
Methodology
a) Materials and apparatus needed
i. Colilert reagent
ii. Sterile transparent 100ml vessel
iii. Temperature controlled incubator
iv. E. coli and coliforms comparator
v. 6W, 365nm Uv lamp
b) Procedure
i. Separate one Colilert pack from its strip
ii. Snap open the reagents pack and pour the reagent in 100ml sample water
iii. Shake to dissolve the reagent
iv. Incubate for 24 hours at 35℃
v. Read the results after 24 hours by comparing the sample to comparator
vi. If the sample is yellow, equal to or greater than the comparator the sample is positive for
total coliforms while if the sample is less yellow than the comparator the sample is
negative for total coliforms.
vii. If the sample is yellow we determine presence of E. coli by shinning Uv light under dark
conditions, shining the light away from your eyes and keeping it within 5 inches of the
vessels.
viii. If the sample is yellow and has fluorescence equal or greater than the comparator, then
the sample is positive for E. coli and vice-versa
ix. If not sure whether the sample is positive incubate for extra 4 hours to allow the
fluorescence to intensify
Quantification of coliforms and E. coli
Quanti-tray system is used for quantification.
There are 2 sizes of Quanti-tray systems i.e.
1. Standard Quality is 51 well that provides count up to 200 per 100 ml sample without
dilution
2. Quanti- tray/ 2000 which has 97 wells and provides count upto 2419 per 100ml sample
without dilution.
Methodology:
Materials and apparatus
i. Quanti-tray x or Quanti-tray x/2000
ii. Quanti- tray comparator for x/2000
iii. Quanti-tray rubber insert
iv. Quanti- tray sealer
v. Number table
Procedure:
i. Turn on the sealer and wait until a green bar appear on the display (Sealer has reached
operating temperature)
ii. Add the Colilert powder in 100ml sample, cap the vessel and shake
iii. Open the quanti- tray by pushing the behind with free hand with the wells facing the arm
iv. Pour the sample mixer into the Quanti- tray (Avoid contact with the foil tab)
v. Place the filled Quanti- tray into the rubber insert making sure each well enters its
corresponding hole
Note: If you need to reverse the motor press and hold the reverse button but don’t reverse the
motor when the rubber insert is inside the sealer
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vi. Remove the rubber insert and tray from the sealer, incubate the sealed tray for 24 hours at
35℃
vii. After incubation mark each yellow with a permanent marker including the large overflow
wells
viii. To determine the number of total coliforms, count the yellow wells that equal to or
greater than the comparator, then refer to MPN table included with the quantities
ix. To determine the of e. coli view the quanti- tray with Uv lamp in a dark environment
shining the light away from your eyes and keep the within 5 inches of the tray
x. Count the fluorescent wells that equal to or greater than the comparator and then refer to
the MPN table
If not sure if the whether the results are positive incubate up to additional 4 hours i.e. 28
hours in total to allow to allow the color or the fluorescence to intensify
Recommendation: Run a quality procedure for each Colilert test.
Summary:
Incubate the sample less yellow than the comparator are negative for for total
coliforms and e. coli
Yellow samples that are equal to or greater than the comparator are positive for the
total coliforms
Yellow and fluorescent samples greater than the comparator are positive for e. coli
