Application of stool-PCR test for diagnosis of Helicobacter pylori infection in children.
OBJECTIVE
To evaluate the usefulness of stool-PCR test for diagnosis of Helicobacter pylori (H pylori) infection in pediatric populations.


METHODS
Based on endoscopic features (including nodular gastritis, erosive duodenitis and ulcer) and/or a positive rapid urease test (RUT) obtained during endoscopy, 28 children from a group of children admitted to the Children's Medical Center of Tehran for persistent upper gastrointestinal problems were selected to compare biopsy-based tests with stool-PCR. Their gastric activity and bacterial density were graded by the updated Sydney system, and their first stool after endoscopy was stored at -70 degrees C. Biopsies were cultured on modified campy-blood agar plates and identified by gram-staining, biochemical tests, and PCR. Two methods of phenol-chloroform and boiling were used for DNA extraction from H pylori isolates. Isolation of DNA from stool was performed using a stool DNA extraction kit (Bioneer Inc, Korea). PCR was performed using primers for detection of vacA, cagA, and 16srRNA genes in both isolates and stool.


RESULTS
Sixteen out of 28 child patients (57%) were classified as H pylori positive by biopsy-based tests, of which 11 (39%) were also positive by stool-PCR. Sensitivity and specificity of stool-PCR was 62.5% and 92.3% respectively. H pylori was observed in histological sections for 10 out of 11 stool-positive patients. Association was observed between higher score of H pylori in histology and positivity of stool-PCR. Also association was observed between the more severe form of gastritis and a positive stool-PCR.


CONCLUSIONS
Association between higher score of H pylori in histology and a positive stool-PCR make it a very useful test for detection of H pylori active infection in children. We also suggest that a simple stool-PCR method can be a useful test for detection of H pylori virulence genes in stool.
Falsafi
Tahereh
T
Department of Biology, Azzahra University, Deh Vanak, 1993891176 Tehran, Iran. tfalsafi@yahoo.com
Favaedi
Raha
R
Mahjoub
Fatemeh
F
Najafi
Mehri
M
D001431:Bacteriological Techniques / Q000379:methods / Q000706:statistics & numerical data; D001483:Base Sequence; D002648:Child; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification; D005243:Feces / Q000382:microbiology*; D016481:Helicobacter Infections / Q000175:diagnosis*; D016480:Helicobacter pylori / Q000235:genetics* / Q000302:isolation & purification; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000706:statistics & numerical data; D012680:Sensitivity and Specificity
D004269:DNA, Bacterial