David Maxwell Lyerly

Fecal Lactoferrin is a sensitive and specific marker in identifying intestinal inflammation

OBJECTIVE:Lactoferrin is a glycoprotein expressed by activated neutrophils. The aim of this study was to determine the sensitivity and specificity of fecal lactoferrin concentrations for inflammatory bowel disease (IBD) or irritable bowel syndrome (IBS) versus healthy controls.METHODS:Fresh stool samples were collected from outpatients with ulcerative colitis (UC), Crohns disease (CD), or IBS. Clinical disease activity for IBD was assessed using a modified Harvey–Bradshaw Activity Index. Fecal lactoferrin concentrations were determined using a polyclonal antibody-based enzyme linked immunoassay. Mean fecal lactoferrin concentrations for each group and sensitivity and specificity of the assay were determined.RESULTS:One hundred-four CD patients, 80 UC patients, 31 IBS patients, and 56 healthy controls were recruited. The mean ± SE fecal lactoferrin concentration (μg/g fecal weight) was 440 ± 128 for CD patients, 1125 ± 498 for UC patients, 1.27 ± 0.29 for IBS patients, and 1.45 ± 0.4 for healthy controls. Fecal lactoferrin was 90% specific for identifying inflammation in patients with active IBD. Elevated fecal lactoferrin was 100% specific in ruling out IBS.CONCLUSIONS:Fecal lactoferrin is sensitive and specific for detecting inflammation in chronic IBD. This noninvasive test may prove useful in screening for inflammation in patients presenting with abdominal pain and diarrhea.

Clostridium difficile Testing: after 20 Years, Still Challenging

More than 20 years ago, as Clostridium difficile was being established as the cause of pseudomembranous colitis and antibiotic-associated diarrhea (AAD), many clinical laboratories were using or beginning to use cycloserine-cefoxitin-fructose agar, a selective medium developed by Lance George and

Fecal Lactoferrin Is a Sensitive and Specific Marker of Disease Activity in Children and Young Adults With Inflammatory Bowel Disease

Background and Aims: Fecal lactoferrin (FLA) is a neutrophil-derived surrogate marker of intestinal inflammation that is elevated in patients with inflammatory bowel disease. However, the correlation between FLA levels and serological markers of disease activity has not been previously reported, to our knowledge. In the present study we evaluated the ability of FLA levels to reflect disease activity in pediatric patients with inflammatory bowel disease. We further assessed the relationship between FLA levels and customary laboratory and clinical measures of inflammation. Patients and Methods: Fecal specimens were collected from 148 consecutive pediatric patients (79 with Crohn disease, 62 with ulcerative colitis, and 7 with irritable bowel syndrome) and 22 healthy control individuals. Lactoferrin was measured by enzyme-linked immunosorbent assay (IBD-SCAN, TECHLAB, Inc). Disease activity was assessed at the time of sample provision by laboratory measures (including erythrocyte sedimentation rate [ESR] and albumin) and previously validated disease activity indices (Pediatric Crohn Disease Activity Index, Kozarek, Harvey Bradshaw Activity Index). Results: Lactoferrin levels were significantly higher in patients with ulcerative colitis (1880 ± 565 μg/mL) (mean ± SE) or Crohn disease (1701 ± 382 μg/mL) than in healthy control individuals under 21 years of age (1.17 ± 0.47 μg/mL, P < 0.001). Lactoferrin levels correlated significantly with ESR, hematocrit, albumin, and platelet count (P < 0.001). Receiver operating characteristic curve analysis revealed that FLA levels were comparable to ESR in detecting patients with clinically active disease (P < 0.001). Patients who experienced a clinical flare within 2 months of specimen collection displayed higher lactoferrin levels (845 ± 452 μg/mL) than did those who remained in clinical remission (190 ± 90 μg/mL, P = 0.003). Conclusions: Data presented here demonstrate that FLA is a sensitive and specific biochemical marker of inflammation for use in the diagnosis and interval assessment of pediatric patients with IBD, and its level correlates well with both clinical disease activity indices and ESR. Elevated levels of FLA may also identify patients at greater risk for the development of subsequent clinical flares.

Fecal lactoferrin: a new parameter to monitor infliximab therapy.

The glycoprotein lactoferrin is found in many body fluids but also in the granules of neutrophilic granulocytes. Fecal lactoferrin levels increase quickly with the influx of leukocytes into the intestinal lumen during inflammation. This biomarker has recently been shown to be a sensitive and specific marker of disease activity in chronic inflammatory bowel disease. Our aim was the determination of fecal lactoferrin as a marker of intestinal inflammation and therapeutic response following infliximab therapy in pediatric patients with Crohns disease (CD). A total of five patients (ages 10-15 years) with severe Crohns disease as defined by the Pediatric Crohns Disease Activity Index (PCDAI) was enrolled in the study. The fecal lactoferrin levels were determined before and after therapy with infliximab by a quantitative lactoferrin ELISA (IBD-SCAN; TechLab, Inc.). Of the five patients on infliximab therapy, three received a single infusion and the remaining two underwent a regime with three maintenance infusions. All five patients responded to infliximab clinically after the first infusion, and in all patients, fecal lactoferrin levels significantly and rapidly decreased from elevated to near baseline in parallel to clinical assessment and the PCDAI. The reduction in fecal lactoferrin at days 7-10 was 93.43 ± 4.49%, in comparison with the level before infliximab therapy, and correlated with a mean decrease in the PCDAI from 48.50 to 14.0. For the patients followed during multiple infusions, one remained with mild disease and the other reached remission (subjective and PCDAI). Fecal lactoferrin is a sensitive and specific biomarker representing intestinal inflammation and response to therapy in pediatric patients with Crohns disease. It may be a helpful noninvasive diagnostic tool for monitoring therapeutic efficiency in pediatric IBD patients. Future studies are needed to further establish the relationship between endoscopic changes and the level of fecal lactoferrin as well as the possible role of lactoferrin as being an early and preclinical indicator of relapse.

Glutamate Dehydrogenase Is Highly Conserved among Clostridium difficile Ribotypes

ABSTRACT gluD was highly conserved and glutamate dehydrogenase (GDH) was readily expressed in vitro by all 77 Clostridium difficile ribotypes assayed. All ribotypes, including ARL 002, ARL 027, and ARL 106, were reactive in assays that detect C. difficile GDH.

Fecal lactoferrin measurements are useful in the interval assessment of patients with active and inactive inflammatory bowel disease

Fecal lactoferrin measurements are useful in the interval assessment of patients with active and inactive inflammatory bowel disease

Clostridia and Bacteroides in Enteric Infections

The ability of anaerobic bacteria to cause disease in humans and animals has been recognized for many years. The clostridia, for example, are well known for their ability to cause disease by producing a variety of toxins ranging from the potent neurotoxins of Clostridium tetani and Clostridium botulinum to the tissue-damaging toxins of the gas gangrene clostridia. Bacteroides fragilis, on the other hand, represents the major anaerobic pathogen in clinical specimens from patients with abscesses and soft-tissue infections. More recently, the importance of members of these genera in gastrointestinal infections has become recognized. Clostridium difficile has emerged as the major cause of nosocomial diarrhea because of its ability to produce diarrhea and pseudomembranous colitis in patients treated with antibiotics. Clostridium perfringens, which is the major cause of gas gangrene, produces a diverse group of toxins that are involved in enteric disease. In industrialized countries, outbreaks of C. perfringens food poisoning occur because of the production of a spore-associated enterotoxin. In certain regions in the Pacific rim, especially in Papua New Guinea, a disease known as pigbel, which is prevalent in persons who eat improperly cooked pork, results from infection with C. perfringens type C strains that produce β-toxin. In addition to these maladies, type E strains of C. perfringens and certain strains of Clostridium spiroforme produce a toxin (iota) that results in another distinctive type of enterotoxemia in animals.

Measurement of anti-saccharomyces cerevisiae antibodies in human feces as a indicator of Crohn’s Disease

Introduction: The method of determining the presence of serum ASCA as a marker of Crohns disease has been used for the differentiation between Crohns disease and ulcerative colitis and has been previously published. In the following study, we describe the first noninvasive method for determining the presence of fecal ASCA. Our approach provides a highly specific method utilizing the extract of Saccharomyces cerevisiae for measuring total fecal ASCA as an aid to distinguish Crohns disease from other gastrointestinal illnesses such as ulcerative colitis and irritable bowel syndrome (IBS). Aim: To describe a novel method for detecting ASCA in human feces and evaluate the clinical utility for distinguishing Crohns disease from ulcerative colitis and IBS. Methods: Fecal ASCA levels were determined qualitatively by an enzyme-linked immunoassay (ASCA-CHEKTM ;TechLab®, Inc.). The immunoassay uses anti-human immunoglobulin antibody conjugated to HRP and microwells coated with Saccharomyces cerevisiae antigens. Fecal specimens were diluted 1:20 in the kit diluent and results were determined by measurement of the optical density (OD) at 450nm/620nm. Results of > 0.200 were considered positive for the presence of fecal ASCA. Typing of immunoglobulins in feces was done using specific human Ig conjugates. Patient Population: Ulcerative colitis (UC): 37 patients comprised of 15 females and 22 males ranging in age from 21-70. Crohns disease (CD): 49 patients comprised of 23 females and 26 males ranging in age from 21-78. Irritable bowel syndrome (IBS): 22 patients comprised of 20 females and 2 males ranging in age 19-78. Healthy control (HC): 12 persons comprised of 8 females and 4 males ranging in age from 20-79. Results: Immunoglobulin Typing of ASCA in Human Fecal Specimens