METHOD FOR DETECTION OF YEAST CELL WALL
Methods are provided for detecting the presence of a yeast cell wall in a food or animal feed suspected of containing the yeast cell wall. The method includes the steps of solubilizing the yeast cell wall, adding a lectin which preferentially binds to a mannose component of the solubilized yeast cell wall, detecting the presence of the lectin. In one embodiment, the lectin preferentially binds to an α-linked mannose component of the yeast cell wall. The α-linked mannose component may be an α-(1,3)-linked mannose or an α-(1,6)-linked mannose, and the yeast cell wall may be derived from Saccharomyces cerevisiae . Kits for accomplishing the method are described, which include a labeled lectin which preferentially binds to a mannose component of the solubilized yeast cell wall and reagents for detecting the labeled lectin.
1 . A method for detecting the presence of a yeast cell wall in a food or animal feed suspected of containing the yeast cell wall, comprising:
solubilizing the yeast cell wall;
adding a lectin which preferentially binds to a mannose component of the solubilized yeast cell wall; and
detecting the presence of the lectin.
2 . The method of claim 1 , wherein the food or animal feed is a grain-based animal feed.
3 . The method of claim 1 , wherein the lectin preferentially binds to an (α-linked mannose component of the yeast cell wall.
4 . The method of claim 3 , wherein the lectin is selected from the group of lectins consisting of Allium ascalonium agglutinin, Allium sativum agglutinin, Allium ursinium agglutinin, Galanthus nivalis agglutinin, Hippeastrum hybrid agglutinin, Helianthus tuberotsus agglutinin, Lens culinaris agglutinin, Listera ovata agglutinin, Narcissus pseudonarcissus lectin, Pisum sativum agglutinin, and combinations thereof.
5 . The method of claim 3 , wherein the lectin preferentially binds to at least one of an α-(1,3)-linked mannose or an α-(1,6)-linked mannose.
6 . The method of claim 5 , wherein the lectin is at least one of a Narcissus pseudonarcissus lectin or a Galanthus nivalis agglutinin.
7 . The method of claim 1 , wherein the solubilized yeast cell wall is bound to a solid surface prior to adding the lectin.
8 . The method of claim 7 , wherein the solubilized yeast cell wall is bound to the solid surface using at least one of sodium chloride or sodium bicarbonate.
9 . The method of claim 5 , wherein the yeast cell wall is derived from Saccharomyces cerevisiae.
10 . A method for detecting the presence of a yeast cell wall contained in a food or animal feed suspected of containing the yeast cell wall, comprising:
solubilizing the yeast cell wall;
adding a lectin which preferentially binds to an α-linked mannose component of the solubilized yeast cell wall; and
detecting the presence of the lectin.
11 . The method of claim 10 , wherein the lectin is selected from the group of lectins consisting of Allium ascalonium agglutinin, Allium sativum agglutinin, Allium ursinium agglutinin, Galanthus nivalis agglutinin, Hippeastrum hybrid agglutinin, Helianthus tuberosus agglutinin, Lens culinaris agglutinin, Listera ovata agglutinin, Narcissus pseudonarcissus lectin, Pisum sativum agglutinin, and combinations thereof.
12 . The method of claim 11 , wherein the lectin preferentially binds to at least one of an α-(1,3)-linked mannose or an α-(1,6)-linked mannose.
13 . The method of claim 12 , wherein the lectin is at least one of a Narcissus pseudonarcissus lectin or a Galanthus nivalis agglutinin.
14 . The method of claim 10 , wherein the solubilized yeast cell wall is bound to a solid substrate prior to adding the lectin.
15 . The method of claim 14 , wherein the solubilized yeast cell wall is bound to the solid surface using at least one of sodium chloride or sodium bicarbonate.
16 . The method of claim 12 , wherein the yeast cell wall is derived from Saccharomyces cerevisiae.
17 . A method for detecting the presence of a yeast cell wall contained in a food or animal feed suspected of containing the yeast cell wall, comprising:
solubilizing the yeast cell wall;
adding a lectin which preferentially binds to an α-(1,3)-linked mannose or an α-(1,6)-linked mannose component of the solubilized yeast cell wall; and
detecting the presence of the lectin.
18 . The method of claim 17 , wherein the lectin is selected from the group of lectins consisting of Allium ascalonium agglutinin, Allium sativum agglutinin, Allium ursinium agglutinin, Galanthus nivalis agglutinin, Hippeastrum hybrid agglutinin, Helianthus tuberosus agglutinin, Lens culinaris agglutinin, Listera ovata agglutinin, Narcissus pseudonarcissus lectin, Pisum sativum agglutinin, and combinations thereof.
19 . The method of claim 18 , wherein the lectin is at least one of a Narcissus pseudonarcissus lectin or a Galanthus nivalis agglutinin.
20 . The method of claim 17 , wherein the solubilized yeast cell wall is bound to a solid surface prior to adding the lectin.
21 . The method of claim 20 , wherein the solubilized yeast cell wall is bound to the solid surface using at least one of sodium chloride or sodium bicarbonate.
22 . The method of claim 19 , wherein the yeast cell wall is derived from Saccharomyces cerevisiae.
23 . A kit for detecting the presence of a yeast cell wall in a food or animal feed in accordance with claim 1 , comprising:
a labeled lectin which preferentially binds to a mannose component of the solubilized yeast cell wall; and
at least one reagent for detecting the labeled lectin.
24 . The kit of claim 23 , wherein the reagent elicits a detectable reaction from the labeled lectin.
25 . The kit of claim 23 , wherein the lectin preferentially binds to an α-(1,3)-linked mannose or an α-(1,6)-linked mannose component of the solubilized yeast cell wall.
26 . The kit of claim 24 , wherein the reaction is detectable by at least one of a calorimetric assay, a fluorescent assay, a chemiluminescent assay, or a radioactive assay.
27 . The kit of claim 26 , wherein the reaction is detectable by a colorimetric assay.
28 . The kit of claim 27 , wherein the reagent further includes a substrate for eliciting a colorimetric reaction from the detector molecule.
29 . The kit of claim 28 , wherein the lectin is labeled with biotin, and the reagent comprises avidin or streptavidin linked to a peroxidase and a chromogenic substrate which elicits a calorimetric reaction from the peroxidase.
30 . The kit of claim 23 , further including a solvent for solubilizing the yeast cell wall.
31 . The kit of claim 30 , wherein the solvent is selected from the group of solvents consisting of dimethyl sulfoxide, sodium hydroxide, potassium hydroxide, boric acid, acetone, water, and combinations thereof.
32 . The kit of claim 31 , wherein the solvent comprises dimethyl sulfoxide.
33 . The kit of claim 25 , wherein the lectin is at least one of a Narcissus pseudonarcissus lectin or a Galanthus nivalis agglutinin.