Cell wall protein CwpV (CD0514) as a diagnostic marker for
Accurate and rapid differentiation of the outbreak strain ribotype 027 from other possible Clostridium difficile ( C. difficile ) strains, using stool samples, facilitates decision making for treatment options. Cell wall protein V (CwpV) contains a cell wall binding domain conserved among C. difficile strains and a variable domain which is antigenically different among C. difficile strains. In embodiments, antibodies against the 027-specific region in CwpV are used in diagnostic tests to detect ribotype 027 in culture or fecal samples.
1. A method of diagnosing a patient with a ribotype 027 C. difficile strain infection that is associated with severe C. difficile infection, the method comprising:
obtaining a fecal sample from a patient;
determining that the patient has a C. difficile infection; and
determining whether the patient has a ribotype 027 C. difficile strain infection by determining a presence of a ribotype 027-specific CwpV antigen in the fecal sample using an immunoassay against the CwpV antigen.
2. The method of claim 1 , wherein the immunoassay is an enzyme-linked immunosorbent assay (ELISA) or an immunochemical reaction on a membrane, or a lateral flow assay.
3. The method of claim 1 , wherein the immunoassay utilizes one or more of monoclonal, polyclonal or recombinant anti-ribotype 027-specific CwpV antibodies as capturing antibodies.
4. The method of claim 1 , wherein the immunoassay utilizes HRP-conjugated polyclonal or monoclonal anti-ribotype 027-specific CwpV antibodies as detection antibodies.
5. The method of claim 1 , wherein the immunoassay comprises one or more antibodies against CwpV raised against native CwpV produced by C. difficile or CwpV derived by recombinant DNA expression in E. coli.
6. The method of claim 1 , wherein determining the presence of the ribotype 027 C. difficile further comprises identifying the presence of a DNA fragment encoding ribotype 027-specific CwpV using nucleic acid amplification assays.
7. The method of claim 1 , further comprising diluting the fecal sample to create a diluted fecal sample; and using the diluted fecal sample in the immunoassay against the CwpV antigen.