Discrepancy between polymerase chain reaction assay and Western blot analysis in the assessment of CagA status in dyspeptic patients.
BACKGROUND
Infection with CagA-positive Helicobacter pylori may be diagnosed by detecting cagA gene by polymerase chain reaction assay (PCR) or serum antibodies against CagA by Western blot analysis. The aim of this study is to evaluate whether results of PCR and Western blot analysis are in agreement in CagA status assessment.


METHODS
Thirty-six dyspeptic patients with unknown H. pylori status underwent upper gastrointestinal endoscopy to assess the presence of mucosal lesions and to collect six gastric biopsies (three from the antrum and three from the body) for evaluation of H. pylori infection (rapid urease test, histology and PCR for ureA gene) and gastritis. CagA status was assessed by PCR (cagA gene) on two biopsy specimens and by Western blot analysis of serum (CagA-antibodies) in each patient.


RESULTS
At endoscopy, nine patients showed normal mucosa, 15 a duodenal ulcer and 12 antral erosions. Twenty-eight patients were found to be H. pylori-positive and eight H. pylori-negative. Of the 28 H. pylori-positive patients, 17 were CagA-positive and five were CagA-negative by both methods, five were CagA-positive by Western blot analysis but not by PCR and one was CagA-positive by PCR but not by Western blot analysis. Of the eight H. pylori-negative patients, none was CagA-positive by PCR, while six were CagA-positive by Western blot analysis. Therefore, the two tests agreed in only 24 patients (67%). In those patients in whom the PCR and Western blot analysis were not in agreement, the histological features appear to suggest that the results of the Western blot analysis should be considered false positives or false negatives.


CONCLUSIONS
PCR and Western blot analysis failed to provide comparable data in many cases. Western blot analysis seems to be more likely to give misleading results than PCR. Thus, PCR seems to be the method of choice to assess CagA status.
Paoluzi
O A
OA
Department of Microbiology, University of Tor Vergata, Italy.
Rossi
P
P
Montesano
C
C
Bernardi
S
S
Carnieri
E
E
Marchione
O P
OP
Nardi
F
F
Iacopini
F
F
Pica
R
R
Paoluzi
P
P
D000328:Adult; D000942:Antigens, Bacterial; D001426:Bacterial Proteins / Q000235:genetics / Q000302:isolation & purification*; D001706:Biopsy; D015153:Blotting, Western; D004386:Duodenum / Q000473:pathology; D004415:Dyspepsia / Q000382:microbiology*; D016145:Endoscopy, Digestive System; D005260:Female; D005753:Gastric Mucosa / Q000473:pathology; D005798:Genes, Bacterial; D016480:Helicobacter pylori / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D007413:Intestinal Mucosa / Q000473:pathology; D008297:Male; D008875:Middle Aged; D016133:Polymerase Chain Reaction; D015203:Reproducibility of Results; D014510:Urease / Q000235:genetics / Q000302:isolation & purification
D000942:Antigens, Bacterial; D001426:Bacterial Proteins; C080866:cagA protein, Helicobacter pylori; D014510:Urease