Rapid coliform detection sytem

A method for determining the quantity of coliform bacteria in a water sample comprising the steps of:

• a) separating said bacteria from said sample using a first filter means;
• b) culturing said bacteria in a broth comprising nutrients for supporting growth of said bacteria and an inducing agent for inducing enzyme production in said bacteria;
• c) separating said bacteria from said broth using a second filter means;
• d) exposing said bacteria to a lysing agent;
• e) incubating a chemiluminogenic substrate of said enzyme with said enzyme to cause cleavage of said substrate, thereby producing a luminescent product;
• f) initiating light emission by exposing said luminescent product to an enhancing agent;
• and,
• g) measuring said light emission to obtain a light measurement corresponding to the quantity of said enzyme to thereby determine the quantity of said bacteria in said sample.

A kit for determining the presence of coliform bacteria in a drinking water sample comprising the steps of:

• a) separating said bacteria from said sample using a first filter means;
• b) culturing said bacteria at a temperature of about 22 to 45° C. for about 2 to 10 hours in a broth comprising nutrients for supporting growth of said bacteria and an inducing agent comprising isopropyl-β-D-thiogalactopyranoside (IPTG) or methyl-β-D-glucuronide (Met-Glu) for inducing production of an enzyme in said bacteria;
• c) separating said bacteria from said broth using a second filter means;
• followed by,
• d) exposing said bacteria on said second filter means to a lysing agent comprising polymyxin-B;
• e) incubating a chemiluminogenic substrate of said enzyme comprising 1,2-dioxetane with said enzyme to cause cleavage of said substrate, thereby producing a luminescent product on said second filter means;
• f) initiating light emission by exposing said luminescent product to an enhancing agent comprising quaternary ammonium homopolymer;
• and,
• g) detecting or measuring said light emission using a luminometer by placing said second filter means with said luminescent product within said luminometer to thereby determine the presence or quantity of said bacteria in said sample.