Cysteine protease Cwp84 (CD2787) as a diagnostic marker for
Accurate and fast detection of the presence of Clostridium difficile ( C. difficile ) disease is crucial for the proper treatment of patients with C. difficile infection. Present tests detecting the presence of C. difficile disease are fast and cost effective, but are not very sensitive. Using an ELISA including Cell Wall Protein 84 (Cwp84) increases the sensitivity of the ELISA. Cwp84 may be used alone or in combination with other markers to support a diagnosis of C. difficile -associated disease.
1. A method for testing a fecal sample from a person, the method comprising:
obtaining the fecal sample from the person;
determining whether Cell Wall Protein 84 (Cwp84) is present in the fecal sample;
upon determining Cwp84 is present in the fecal sample, concluding the presence of C. difficile in the fecal sample; and
diagnosing the person with active C. difficile infection upon determining the presence of C. difficile in the fecal sample.
2. The method of claim 1 , wherein the presence of C. difficile may be further concluded based upon the presence of glutamate dehydrogenase (GDH) and toxins A and B in the fecal sample.
3. The method of claim 1 , wherein the presence of Cwp84 is identified using enzyme-linked immunosorbent assays.
4. The method of claim 3 , wherein one or more antibodies against Cwp84 are raised against a native Cwp84 purified from C. difficile strains.
5. The method of claim 3 , wherein one or more antibodies against Cwp84 are raised against Cwp84 recombinantly expressed in E. coli.
6. The method of claim 1 , wherein the presence of Cwp84 is identified using lateral flow techniques and immunochemical reactions on a membrane.
7. The method of claim 6 , wherein one or more antibodies against Cwp 84 are raised against a native Cwp84 purified from C. difficile strains.
8. The method of claim 6 , wherein one or more antibodies against Cwp84 are raised against Cwp84 recombinantly expressed in E. coli .
9. The method of claim 3 , wherein one or more antibodies against Cwp84 are raised against a full length sequence of Cwp84.
10. The method of claim 3 , wherein one or more antibodies against Cwp84 are raised against one or more variants of Cwp84 from one or more C. difficile isolates.
11. The method of claim 3 , wherein one or more antibodies against Cwp84 are raised against one of active Cwp84 or inactive Cwp84 containing one or more mutations.
12. The method of claim 11 , wherein the one or more mutations include C116A.
13. The method of claim 2 , wherein the presence of GDH and Toxins A and B is measured using one or more of enzyme-linked immunoassays, lateral flow, and immunochemical reactions on a membrane.
14. A method for testing a fecal sample from a person, the method comprising:
obtaining the fecal sample from the person;
determining whether C. difficile is present in the fecal sample based on the presence of Cell Wall Protein 84 (Cwp84) and glutamate dehydrogenase (GDH);
upon determining the presence of either GDH or Cwp84 in the fecal sample, concluding presence of C. difficile in the fecal sample; and
diagnosing the person with active C. difficile infection upon determining the presence of C. difficile in the fecal sample.
15. The method of claim 14 , further comprising identifying the presence of Toxin A and Toxin B in the fecal sample, wherein the presence of Cwp84, GDH, and Toxins A and B is identified utilizing at least one of enzyme-linked immunoassays, lateral flow, and immunochemical reactions on a membrane.
16. The method of claim 15 , wherein one or more antibodies against Cwp84 are raised against one of a full length sequence of Cwp84.
17. The method of claim 15 , wherein one or more antibodies against Cwp84 are raised against one of active Cwp84 or inactive Cwp84 containing a C116A mutation.