Method for detecting microorganisms
View Patent ↗Described herein are methods of detecting a wound infection and for detecting the presence or absence of microorganisms, for example, wound pathogens in a sample, by contacting a sample with an enzyme produced and/or secreted by the bacteria, and detecting modification or the absence of modification of the substrate, as an indicator of the presence or absence of the enzyme in the sample. The present invention also features a biosensor for detecting the presence or absence of bacteria in a sample.
1. A method for detecting the presence or absence of a wound pathogenic bacterium in a sample of wound fluid obtained from a subject, comprising the steps of:
(a) contacting the sample with a detectably labeled polypeptide substrate comprising the amino acid sequence of SEQ ID NO: 1, 2, 3, 4, 5, 9 or 10, or an amino acid sequence having at least 99% sequence identity to SEQ ID NO: 1, 2, 3, 4, 5, 9 or 10; and
(b) detecting the modification or the absence of the modification of said substrate wherein the presence of said modification of said substrate indicates the presence of said wound-pathogenic bacterium in said sample, and the absence of modification of said substrate indicates the absence of said wound-pathogenic bacterium in said sample.
2. The method of claim 1 , wherein said substrate is on a solid support.
3. The method of claim 2 , wherein said solid support comprises a material required to be free of microbial contaminants.
4. The method of claim 2 , wherein said solid support is selected from the group consisting of a wound dressing, a container for holding body fluids, a disk, a scope, a filter, a lens, foam, cloth, paper, a suture, and a swab.
5. The method of claim 4 , wherein said container for holding body fluids is selected from the group consisting of a urine collection bag, a blood collection bag, a plasma collection bag, a test tube, a catheter, and a well of a microplate.
6. A method for detecting the presence or absence of Staphylococcus aureus in a sample of wound fluid obtained from a subject, comprising the steps of:
(a) contacting the sample with a detectably labeled polypeptide substrate comprising the amino acid sequence of SEQ ID NO: 1 or having at least 95% sequence identity to SEQ ID NO: 1, the substrate being for a protease produced and/or secreted by said Staphylococcus aureus but not produced and/or secreted by a non-wound-pathogenic bacterium or the subject, under conditions that result in modification of said substrate by said protease; and
(b) detecting the modification or the absence of the modification of said substrate, wherein modification of said substrate indicates the presence of Staphylococcus aureus in said sample, and wherein the absence of modification of said substrate indicates the absence of Staphylococcus aureus in said sample.
7. The method of claim 6 , wherein said substrate is on a solid support.
8. The method of claim 7 , wherein said solid support comprises a material required to be free of microbial contaminants.
9. The method of claim 7 , wherein said solid support is selected from the group consisting of a wound dressing, a container for holding body fluids, a disk, a scope, a filter, a lens, foam, cloth, paper, a suture, and a swab.
10. The method of claim 9 , wherein said container for holding body fluids is selected from the group consisting of a urine collection bag, a blood collection bag, a plasma collection bag, a test tube, a catheter, and a well of a microplate.