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Inflammatory bowel disease
Crohn’s disease and Ulcerative
colitis
By Sakar Ahmed Abdullah
Sakar.Ahmed@spu.edu.iq
Technical College of Health
Medical Laboratory Department
Outline
• Inflammatory bowel disease
• Diagnostic criteria
• Materials
• Method
• Result interpretation
Inflammatory bowel disease
• •Inflammatory bowel disease (IBD) is a chronic inflammatory disorder of the
gastrointestinal tract includs ulcerative colitis and Crohn's disease with courses of
relapse and remission.
• Ulcerative colitis (UC) characterized by mucosal ulceration that appears in a
continuous pattern. Ulcerative colitis only affects the large intestine.
• Crohn's disease may affect any part of the digestive system from the mouth to the
anus most commonly occurring at the end of the small intestine, appearing in
patches and extending through the entire thickness of the bowel wall (transmural).
Diagnostic criteria
1. Physicians depend on symptoms
• Bloody diarrhea
• Mucus in the stool
• Abdominal cramping, flatulence
• Fatigue, weight loss, malnutrition, anemia
• The untreated case may cause colon cancer
• Toxic megacolon
• Mouth ulcers (Crohn’s disease )
• Perianal lesions and bleeding (Crohn’s disease )
2. Biopsy
• The biopsy will take by colonoscopy for diagnosing the disease stages in
ulcerative colitis. The specimen should take from the most severe parts. The
active disease could be diagnosed by the presence of neutrophils, damaged
epithelial cells, irregular surface, and crypt abnormalities.
• In Crohn's disease, fissures will be observed by endoscopy especially in the small
intestine.
3. Laboratory investigation
1. CBC for detecting (anemia, increasing WBC and platelet)
2. Biochemical tests (iron, albumin, creatinine).
3. Inflammatory biomarker tests such as CRP, ESR.
4. liver function tests
• Blood Urea Nitrogen (BUN) decrease
• Aspartate Aminotransferase (AST) increase
• Alanine Aminotransferase (ALT) increase
• Alkaline Phosphatase (ALP) increase
• Gamma-glutamyl Transpeptidase (GGT) increase
• Total Bilirubin (T.Bil), Direct Bilirubin(D.Bil) increase
5. Microbiological tests and culturing stool sample to differentiate colitis from other infectious pathogens.
6. Cytokines such Tumor necrosis factor (TNF), interleukin 1 and interleukin6 are elevated.
7. T helper cell count (T helper cell 1 elevated in Crohn's disease while T helper cell 2 elevated in ulcerative colitis)
detect by flowcytometry.
8. Bloody diarrhea detect by fecal occult blood test which detect Hb (using immunochromatographic assay )
9. Fecal calprotectin test will be positive.
Fecal calprotectin (calfast)
• Calprotectin is a protein has bacteriostatic and fungistatic properties released by a neutrophil.
Elevated faecal calprotectin indicates the migration of neutrophils to the intestinal mucosa.
• Fecal calprotectin test is the measurement of the protein calprotectin in the stool that is used to
1. Distinguish between IBD and non-inflammatory disorders
2. Detecting activity and severity of inflammatory bowel disease.
4. Serological test for detecting autoantibodies by
1. IgG and IgA "perinuclear Anti-neutrophil cytoplasmic antibodies (p-ANCA) in
ulcerative colitis
2. Anti-Saccharomyces cerevisiae antibodies (ASCA) in crohn’s disease
3. Anti-Calprotectin Antibody
4. Anti lactoferrin antibody
5. Anti myeloperoxidase antibody
Fecal calprotectin test by immunochromatographic assay
• The liquid sample is dropped on the well, the antigen in the sample
produces a complex with free antibody labeled with colloidal gold.
• Then complex moves along with the liquid sample on the
nitrocellulose membrane bind with the bound antibody, producing
a red-purple line that indicates the presence of antigen in
the sample.
• Labeled antibody move through the membrane and bind with
second immobilized antibody producing purple color in control line
Method
Method
1. Collect a stool sample in a sterile container
2. Extract the device shaped-rod unscrewing the cap counter-clockwise.
3. Dip the rod shaped section into the stool sample.
4. Rotate the rod in the sample several times until all grooves have been
filled with stool.
5. insert the shaped rod with the faecal material inside the tube containing
the extraction fluid and rotate the cap clockwise until completely closed.
6. Dispense 2 drops of the sample to the well of the cassette, do not
overload the sample
7. Result can be read after 10 min
Reference
• https://discovery.wehi.edu.au/timeline/neuraminidase
• http://www.antibodies-
online.com/kit/1305182/Fecal+Calprotectin+Rapid+Test+Kit/
• https://link.springer.com/chapter/10.1007%2F978-1-4757-9182-2_97

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lab3 inflammatory bowel disease-chromatography .pdf

  • 1. Inflammatory bowel disease Crohn’s disease and Ulcerative colitis By Sakar Ahmed Abdullah Sakar.Ahmed@spu.edu.iq Technical College of Health Medical Laboratory Department
  • 2. Outline • Inflammatory bowel disease • Diagnostic criteria • Materials • Method • Result interpretation
  • 3. Inflammatory bowel disease • •Inflammatory bowel disease (IBD) is a chronic inflammatory disorder of the gastrointestinal tract includs ulcerative colitis and Crohn's disease with courses of relapse and remission. • Ulcerative colitis (UC) characterized by mucosal ulceration that appears in a continuous pattern. Ulcerative colitis only affects the large intestine. • Crohn's disease may affect any part of the digestive system from the mouth to the anus most commonly occurring at the end of the small intestine, appearing in patches and extending through the entire thickness of the bowel wall (transmural).
  • 4.
  • 5. Diagnostic criteria 1. Physicians depend on symptoms • Bloody diarrhea • Mucus in the stool • Abdominal cramping, flatulence • Fatigue, weight loss, malnutrition, anemia • The untreated case may cause colon cancer • Toxic megacolon • Mouth ulcers (Crohn’s disease ) • Perianal lesions and bleeding (Crohn’s disease )
  • 6. 2. Biopsy • The biopsy will take by colonoscopy for diagnosing the disease stages in ulcerative colitis. The specimen should take from the most severe parts. The active disease could be diagnosed by the presence of neutrophils, damaged epithelial cells, irregular surface, and crypt abnormalities. • In Crohn's disease, fissures will be observed by endoscopy especially in the small intestine.
  • 7.
  • 8. 3. Laboratory investigation 1. CBC for detecting (anemia, increasing WBC and platelet) 2. Biochemical tests (iron, albumin, creatinine). 3. Inflammatory biomarker tests such as CRP, ESR. 4. liver function tests • Blood Urea Nitrogen (BUN) decrease • Aspartate Aminotransferase (AST) increase • Alanine Aminotransferase (ALT) increase • Alkaline Phosphatase (ALP) increase • Gamma-glutamyl Transpeptidase (GGT) increase • Total Bilirubin (T.Bil), Direct Bilirubin(D.Bil) increase 5. Microbiological tests and culturing stool sample to differentiate colitis from other infectious pathogens. 6. Cytokines such Tumor necrosis factor (TNF), interleukin 1 and interleukin6 are elevated. 7. T helper cell count (T helper cell 1 elevated in Crohn's disease while T helper cell 2 elevated in ulcerative colitis) detect by flowcytometry. 8. Bloody diarrhea detect by fecal occult blood test which detect Hb (using immunochromatographic assay ) 9. Fecal calprotectin test will be positive.
  • 9. Fecal calprotectin (calfast) • Calprotectin is a protein has bacteriostatic and fungistatic properties released by a neutrophil. Elevated faecal calprotectin indicates the migration of neutrophils to the intestinal mucosa. • Fecal calprotectin test is the measurement of the protein calprotectin in the stool that is used to 1. Distinguish between IBD and non-inflammatory disorders 2. Detecting activity and severity of inflammatory bowel disease.
  • 10. 4. Serological test for detecting autoantibodies by 1. IgG and IgA "perinuclear Anti-neutrophil cytoplasmic antibodies (p-ANCA) in ulcerative colitis 2. Anti-Saccharomyces cerevisiae antibodies (ASCA) in crohn’s disease 3. Anti-Calprotectin Antibody 4. Anti lactoferrin antibody 5. Anti myeloperoxidase antibody
  • 11. Fecal calprotectin test by immunochromatographic assay • The liquid sample is dropped on the well, the antigen in the sample produces a complex with free antibody labeled with colloidal gold. • Then complex moves along with the liquid sample on the nitrocellulose membrane bind with the bound antibody, producing a red-purple line that indicates the presence of antigen in the sample. • Labeled antibody move through the membrane and bind with second immobilized antibody producing purple color in control line
  • 13. Method 1. Collect a stool sample in a sterile container 2. Extract the device shaped-rod unscrewing the cap counter-clockwise. 3. Dip the rod shaped section into the stool sample. 4. Rotate the rod in the sample several times until all grooves have been filled with stool. 5. insert the shaped rod with the faecal material inside the tube containing the extraction fluid and rotate the cap clockwise until completely closed. 6. Dispense 2 drops of the sample to the well of the cassette, do not overload the sample 7. Result can be read after 10 min