UreC PCR based diagnosis of Helicobacter pylori infection and detection of cag A gene in gastric biopsies.
Polymerase chain reaction assay using ureC gene specific primers for the detection of Helicobacter pylori in gastric biopsy specimens from 116 dyspeptic patients was compared with other routine invasive diagnostic methods (culture, rapid urease test [RUT] and histology). In parallel, gastric biospy specimens from 54 patients and their corresponding Helicobacter pylori isolates were subjected to PCR with cagA targeting primers using standard protocols. Helicobacter pylori were detected in 53%, 43%, 48% and 50% of patients by PCR, RUT, culture and histological examination respectively. Based on histology and culture positive and at least three test positive result, 44 (37%), 46 (39%) and 26 (22%), and 56 (48%), 52 (44%) and 8 (6%) patients were classified as Helicobacter pylori positive, negative and indeterminate respectively. The sensitivity and specificity of PCR assay was the highest-95% and 100% when compared with both culture and histology positive, and at least any three positive results respectively. The result of cagA positivity in 54 gastric biopsy specimens and their corresponding Helicobacter pylori isolates were identical; 18 of 20 (90%) duodenal ulcer patients and 23 of 28 (82%) patients with chronic gastritis and 2 (40%) of 5 patients with portal hypertension and one gastric biopsy specimens from gastric cancer patients were found to be cagA positive. PCR-based method to detect Helicobacter pylori and the virulence gene cag A directly from gastric biopsy specimens appears to be promising and can curtail the lengthy process of culture-based approaches. The procedure proved to be rapid and reliable and could be utilized for diagnostic purposes.
Mishra
K K
KK
Department of Microbiology, Sanjay Gandhi Postgraduate Institute of Medical Science, Lucknow, India.
Srivastava
S
S
Dwivedi
P P
PP
Prasad
K N
KN
Ayyagari
A
A
D000328:Adult; D000368:Aged; D000942:Antigens, Bacterial; D001426:Bacterial Proteins / Q000235:genetics*; D001706:Biopsy; D003470:Culture Media; D004415:Dyspepsia; D005260:Female; D005798:Genes, Bacterial; D016481:Helicobacter Infections / Q000175:diagnosis* / Q000382:microbiology / Q000473:pathology; D016480:Helicobacter pylori / Q000235:genetics / Q000302:isolation & purification; D006801:Humans; D008297:Male; D008875:Middle Aged; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013270:Stomach / Q000382:microbiology* / Q000473:pathology; D014510:Urease / Q000235:genetics*
D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D003470:Culture Media; C080866:cagA protein, Helicobacter pylori; D014510:Urease