APT Test (Fetal Hemoglobin)

Blood in the gastrointestinal tract or stool of neonates - Swallowed blood of maternal origin or secondary to disease Qualitative test - determination in grossly bloody stools or hematemesis from a newborn.

APT Test Procedure

1. Material to be tested is emulsified in water to release hemoglobin 2. Centrifuge 3. The pink supernatant is mixed with 1% sodium hydroxide in a ratio of 5:1 4. Wait for 2 minutes then compare with control tubes of maternal and fetal hemoglobin

Fetal hemoglobin Hb F Alkali-resistant Solution remains pink Maternal hemoglobin Hb A Denatured Yellow-brown supernatant Also distinguishes between maternal hemoglobins AS, CS, and SS, and fetal hemoglobin Presence of thalassemia major erroneous result due to hemoglobin F Stool should be tested when fresh

Fecal Enzymes
Production of enzymes - chronic pancreatitis and cystic fibrosis steatorrhea and presence of undigested food in the feces Focused primarily on proteolytic enzymes: 1. Trypsin 2. Chymotrypsin 3. Elastase I

Trypsin
Central enzyme involved in activation of all digestive proenzymes Absence is screened by exposing x-ray paper to stool emulsified in water Inability to digest the gelatin on the paper indicates deficiency in production Detects only severe cases of pancreatic insufficiency False (-) results intestinal degradation and trypsin inhibitors in feces False (+) results bacterial enzymes in old specimens

Chymotrypsin
More resistant to intestinal degradation More sensitive indicator of less severe cases of pancreatic insufficiency Stable for up to 10 days at room temperature Most frequently measured by spectrophotometry

Elastase I
Elastase isoenzyme from pancreas Concentrations in pancreatic secretions (6% of all secreted pancreated enzymes) Strongly resistant to degradation Concentration in pancreas is 5X > than in pancreatic juice Not affected by motility disorders or mucosal defects Measured through ELISA ELISA test uses monoclonal antibodies against human pancreatic elastase-1 Specific in differentiating pancreatic from non pancreatic causes in patients with steatorrhea

Carbohydrates
Osmotic diarrhea presence of increased amounts draws in fluids and electrolytes Due to celiac disease or lactose intolerance Primarily analyzed by serum and urine tests, Concentration Copper Reduction test Tests for fecal reducing substances - congenital disaccharide and enzyme deficiencies Infant diarrhea Normal stool pH: 7-8, < 5.5 indicates intestinal bacterial fermentation and carbohydrate disorders

Clinitest
Copper Reduction test 0.5 g/dL Indicative of carbohydrate intolerance Diarrhea caused by abnormal excretion of reducing sugars/Viral or parasitic etiology Sucrose not detected Premature infants: Correlation between (+) result and inflammatory necrotizing entercolitis General test for reducing sugars, (+) result followed by Dxylose test for malabsorption and lactose tolerance test for maldigestion Small bowel biopsy and assay of disaccharidase enzyme activity

D-Xylose Test
Differential diagnosis of malabsorption A 25-g dose of pentose sugar in water is administered orally, and the amount excreted over a 5-hour period in the urine is determined

< 3 g Enterogenous malabsorption (Pancreatic enzymes are not required for absorption of D-xylose) D-xylose is passively absorbed in the small intestine and not metabolized by the liver Accuracy depends on the rate of absorption and on the rate of excretion.

Lactose tolerance test

Following an overnight fast, administer orally 50 g of lactose dissolved in 400 mL of water. Draw fasting blood and blood samples at 30, 60, and 120 minutes after ingestion (Glucose tolerance test). Patients with lactase deficiency exhibit a peak rise of less than 20 mg/dL