Detection of Helicobacter pylori gene expression in human gastric mucosa.
Mucosal and systemic immunologic recognition of cagA by Helicobacter pylori-infected individuals is associated with peptic ulcer disease; however, in the laboratory, expression of cagA is subject to artificial conditions which may not accurately reflect the conditions in host tissues. Gastric antral and body biopsy specimens and serum for anti-H. pylori immunoglobulin G serology were obtained from 42 patients. Biopsy specimens were studied by histology, culture, and reverse transcription PCR (RT-PCR). Oligonucleotide primers specific for H. pylori (16S rRNA, ureA, and cagA) were used to detect bacterial mRNA in gastric biopsy specimens. PCR was performed on DNA from corresponding H. pylori isolates to detect genomic 16S rRNA, ureA, and cagA. Of the 42 patients from whom clinical specimens were obtained, 25 were infected with H. pylori on the basis of both serology and histology or culture (i.e., tissue positive); 13 were negative by serology, histology, and culture; and 4 were positive by serology only. RT-PCR with 16S rRNA primers detected 24 of 25 tissue-positive and 0 of 17 tissue-negative patients (P < 0.001). RT-PCR with ureA primers detected 16 of 25 tissue-positive and 0 of 17 tissue-negative patients (P < 0.001). CagA mRNA was detected by RT-PCR in 14 of 25 gastric biopsy specimens in the tissue-positive group and in 0 of 17 gastric biopsy specimens in the tissue-negative group. PCR of genomic DNA for the presence of the cagA gene in the corresponding bacterial isolates correlated absolutely with cagA gene expression in gastric tissue. These results indicate that RT-PCR is a sensitive and specific method for the detection of the presence of H. pylori and the expression of H. pylori genes in human gastric tissue. Detection of H. pylori gene expression in vivo by this approach may contribute to improving the diagnosis and understanding the pathogenesis of H. pylori infections.
Peek
R M
RM
Department of Medicine, Vanderbilt University School of Medicine, Nashville, Tennessee 37232-2279.
Miller
G G
GG
Tham
K T
KT
Pérez-Pérez
G I
GI
Cover
T L
TL
Atherton
J C
JC
Dunn
G D
GD
Blaser
M J
MJ
D000942:Antigens, Bacterial; D001426:Bacterial Proteins / Q000235:genetics / Q000302:isolation & purification; D001483:Base Sequence; D001706:Biopsy; D017931:DNA Primers; D005753:Gastric Mucosa / Q000382:microbiology*; D016481:Helicobacter Infections / Q000175:diagnosis* / Q000235:genetics / Q000276:immunology; D016480:Helicobacter pylori / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods; D012336:RNA, Ribosomal, 16S / Q000235:genetics / Q000302:isolation & purification; D014510:Urease / Q000235:genetics / Q000302:isolation & purification
D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D017931:DNA Primers; D012336:RNA, Ribosomal, 16S; C080866:cagA protein, Helicobacter pylori; D014510:Urease