Fecal lactoferrin as a biomarker for determining disease severity and for monitoring infection in patients with clostridium difficile disease

Abstract

Clostridium difficile disease involves a range of clinical presentations ranging from mild to self-limiting diarrhea to life-threatening pseudomembranous colitis and megacolon. Cases of C. difficile are treated differently depending on severity of disease. Mild and moderate cases may be treated with metronidazole while moderate-to-severe and relapsing cases are often treated with vancomycin or fidaxomicin. The presence of C. difficile disease is detected using a biomarker panel that includes C. difficile antigen (GDH), toxins A and B, and fecal lactoferrin. In patients suspected of C. difficile disease, if GDH is detected indicating the presence of C. difficile , and then toxins A and/or B are detected to indicate toxigenic C. difficile and support a diagnosis of C. difficile -associated disease, fecal lactoferrin concentrations are measured to determine severity of the disease by indicating the amount of intestinal inflammation.

1 . A method of monitoring a patient with  C. Difficile  disease, the method comprising:
 obtaining a first fecal sample from a patient at a first time;   obtaining a second fecal sample from the same patient at a second time later than the first time;   comparing a first amount of one or more of lactoferrin or calprotectin in the first fecal sample with a second amount of one or more of lactoferrin or calprotectin in the second fecal sample to identify a change in level of one or more of lactoferrin or calprotectin between the first time and the second time;   based on a change in level of one or more of lactoferrin or calprotectin, identifying a patient's change in  C. Difficile  disease severity; and   administering a therapeutically effective amount of a treatment shown to be effective in treating  C. Difficile  to the patient based on identifying a patient's change in  C. Difficile  disease severity.   
     
     
         2 . The method of  claim 1 , wherein a therapeutically effective amount of the treatment is stopping a treatment if the level of lactoferrin has dropped below 7.25 μg/mL for the second fecal sample of the patient. 
     
     
         3 . The method of  claim 1 , wherein a therapeutically effective amount of the treatment is administering a therapeutically effective amount of a treatment shown to be effective in treating moderate  C. Difficile  if the level of lactoferrin is between 7.25 μg/mL and 99.99 μg/mL for the second fecal sample of the patient. 
     
     
         4 . The method of  claim 3 , wherein the treatment shown to be effective in treating moderate  C. Difficile  comprises one or more of nitroimidazole antibiotics. 
     
     
         5 . The method of  claim 1 , wherein a therapeutically effective amount of the treatment is administering a therapeutically effective amount of a treatment shown to be effective in treating moderate-to-severe  C. Difficile  if the level of lactoferrin is 100 μg/mL or greater for the second fecal sample of the patient, wherein a level of lactoferrin of 100 μg/mL or greater indicates severe intestinal inflammation. 
     
     
         6 . The method of  claim 5 , wherein treatment shown to be effective in treating moderate-to-severe  C. Difficile  comprises treatment with one or more of glycopeptide antibiotics or macrocyclic antibiotics. 
     
     
         7 . The method of  claim 5 , wherein treatment shown to be effective in treating moderate-to-severe  C. Difficile  comprises treatment with a native flora transplant. 
     
     
         8 . A method of monitoring a patient with  C. Difficile  disease, the method comprising:
 obtaining a first fecal sample from a patient at a first time;   obtaining a second fecal sample from the same patient at a second time later than the first time;   comparing a first amount of one or more of lactoferrin or calprotectin in the first fecal sample with a second amount of one or more of lactoferrin or calprotectin in the second fecal sample to identify a change in level of one or more of lactoferrin or calprotectin between the first time and the second time;   based on the change in level of one or more of lactoferrin or calprotectin, identifying a patient's change in  C. Difficile  disease severity; and   administering a therapeutically effective amount of a treatment shown to be effective in treating  C. Difficile  to the patient after obtaining the second fecal sample, wherein the patient had a mild case of  C. Difficile  at the first time and an increased amount of one or more of lactoferrin or calprotectin at the second time, wherein the comparison of the first amount in the first sample and the second amount in the second sample indicates increased intestinal inflammation and worsening of the  C. Difficile  disease.   
     
     
         9 . The method of  claim 8 , wherein a therapeutically effective amount of the treatment is administering a therapeutically effective amount of a treatment shown to be effective in treating moderate  C. Difficile  if the level of lactoferrin is between 7.25 μg/mL and 99.99 μg/mL for the second fecal sample of the patient. 
     
     
         10 . The method of  claim 9 , wherein the treatment shown to be effective in treating moderate  C. Difficile  comprises treatment with one or more of nitroimidazole antibiotics. 
     
     
         11 . The method of  claim 8 , wherein a therapeutically effective amount of the treatment is administering a therapeutically effective amount of the treatment shown to be effective in treating moderate-to-severe  C. Difficile  if the level of lactoferrin is 100 μg/mL or greater for the second fecal sample of the patient. 
     
     
         12 . The method of  claim 11 , wherein the treatment shown to be effective in treating moderate-to-severe  C. Difficile  comprises treatment with one or more of glycopeptide antibiotics or macrocyclic antibiotics. 
     
     
         13 . The method of  claim 11 , wherein the wherein the treatment shown to be effective in treating moderate-to-severe  C. Difficile  comprises treatment with a native flora transplant. 
     
     
         14 . A diagnostic method of determining a presence of  C. Difficile  disease and a severity of  C. Difficile  disease, the method comprising:
 obtaining a fecal sample from a patient;   determining a presence of a first biomarker from the same patient's fecal sample, wherein the presence of the first biomarker indicates the presence of  C. Difficile  disease; and   determining a level of a second biomarker from the same patient's fecal sample, wherein the level of the second biomarker indicates the severity of the  C. Difficile  disease.   
     
     
         15 . The method of  claim 14 , wherein the first biomarker comprises  C. difficile  glutamate dehydrogenase (GDH). 
     
     
         16 . The method of  claim 14 , wherein the first biomarker comprises  C. Difficile  toxin A. 
     
     
         17 . The method of  claim 14 , wherein the first biomarker comprises  C. Difficile  toxin B. 
     
     
         18 . The method of  claim 14 , wherein the first biomarker comprises one or more of  C. Difficile  toxin A gene (tcdA) or  C. Difficile  toxin B gene (tcdB). 
     
     
         19 . The method of  claim 14 , wherein the second biomarker comprises a level of lactoferrin in the patient's fecal sample. 
     
     
         20 . The method of  claim 14 , wherein the second biomarker comprises a level of calprotectin in the patient's fecal sample.