Composition for detecting peptidoglycan, and diagnostic kit detecting peptidoglycan

Abstract

The present invention relates to a composition for selectively detecting an extremely small amount of peptidoglycan in sample, a preparation method of the composition, and a detection kit for peptidoglycan. It is possible to quantify a small amount of peptidoglycan contained in human blood, tissue, body fluid, water or food, and to diagnose an infection of microorganism with peptidoglycan as a component of cell wall using the composition and the detection kit. In addition, the composition can be applied for a diagnosis reagent of detecting an infection of Gram-positive bacteria in animal or human being in advance, and thus, can be used for the prevention or treatment of food poisonings and Bacterial sepsis.

1 . A composition for detecting a peptidoglycan comprising extract of insect body fluid which has a phenoloxidase activity on the peptidoglycan without the addition of calcium.  
     
     
         2 . The composition according to  claim 1 , wherein the extract of insect body fluid is a plasma solution separated from insect body fluid.  
     
     
         3 . The composition according to  claim 1 , wherein the extract of insect body fluid comprises a plasma solution and hemocyte lysate of insect body fluid.  
     
     
         4 . The composition according to  claim 1 , wherein the extract of insect body fluid is derived from  Galleria mellanella  larvae.  
     
     
         5 . The composition according to  claim 2 , wherein the extract of insect body fluid is a fraction having a phenoloxidase activity on peptidoglycan without the addition of calcium selected from fractions which are prepared by treating plasma of insect body fluid with solvent or buffer solution.  
     
     
         6 . The composition according to  claim 3 , wherein the extract of insect body fluid is a fraction having a phenoloxidase activity on peptidoglycan without the addition of calcium selected from fractions which are prepared by lysing hemocyte contained in insect body fluid containing plasma and hemocyte, and treating with solvent or buffer solution.  
     
     
         7 . The composition according to  claim 3 , wherein the extract of insect body fluid is a fraction having a phenoloxidase activity on peptidoglycan without the addition of calcium selected from fractions which are prepared by adding hemocyte lysate or partially-purified hemocyte lysate to fractions obtained by treating plasma of  Galleria mellanella  larvae with solvent or buffer solution.  
     
     
         8 . The composition according to  claim 5 , wherein the solvent or buffer solution comprises a chelating agent in a sufficient amount for chelating calcium ion existing in a sample and separation process.  
     
     
         9 . The composition according to  claim 5 , wherein the fraction is a fraction prepared through column chromatography.  
     
     
         10 . The composition according to  claim 9 , wherein the column is filed with a sugar resin or a vinyl resin.  
     
     
         11 . A detection method of peptidoglycan comprising the steps of: obtaining of sample from the test subject, adding a composition for detecting peptidoglycan comprising extract of insect body fluid which has a phenoloxidase activity on the peptidoglycan without the addition of calcium to the sample, and measuring a phenoloxidase activity.  
     
     
         12 . The detection method according to  claim 11 , wherein the extract of insect body fluid is a fraction having a phenoloxidase activity on peptidoglycan without the addition of calcium selected from fractions which are prepared by treating plasma of insect body fluid with solvent or buffer solution.  
     
     
         13 . The detection method according to  claim 11 , wherein the extract of insect body fluid is a fraction having a phenoloxidase activity on peptidoglycan without the addition of calcium selected from fractions which are prepared by lysing hemocyte in insect body fluid containing plasma and hemocyte, and treating with solvent or buffer solution.  
     
     
         14 . The detection method according to  claim 11 , wherein the extract of insect body fluid is a fraction having a phenoloxidase activity on peptidoglycan without the addition of calcium selected from fractions which are prepared by adding hemocyte lysate or partially-purified hemocyte lysate to fractions obtained by treating plasma of  Galleria mellonella  larvae with solvent or buffer solution.  
     
     
         15 . A detection kit for peptidoglycan comprising the composition for detecting a peptidoglycan comprising extract of insect body fluid which has a phenoloxidase activity on the peptidoglycan without the addition of calcium.  
     
     
         16 . The detection kit according to  claim 15 , wherein the extract of insect body fluid is a fraction having a phenoloxidase activity on peptidoglycan without the addition of calcium selected from fractions which are prepared by treating plasma of insect body fluid with solvent or buffer solution.  
     
     
         17 . The detection kit according to  claim 15 , wherein the extract of insect body fluid is a fraction having a phenoloxidase activity on peptidoglycan without the addition of calcium selected from fractions which are prepared by lysing hemocyte in insect body fluid containing plasma and hemocyte, and treating with solvent or buffer solution.  
     
     
         18 . The detection kit according to  claim 15 , wherein the extract of insect body fluid is a fraction having a phenoloxidase activity on peptidoglycan without the addition of calcium selected from fractions which are obtained by adding hemocyte lysate or partially-purified hemocyte lysate to fractions obtained by treating plasma of  Galleria mellonella  larvae with solvent or buffer solution.