[Development of a detection system for Helicobacter pylori DNA in gastric juice].
A rapid and sensitive PCR-based microwell plate assay (PCR-MWP) system to detect the 16 S ribosome RNA gene of Helicobacter pylori was developed. Analytical sensitivity, evaluated with purified recombinant plasmid DNA and genomic DNA of H. pylori, was one copy of DNA per PCR. Specificity was validated with a panel of DNA from 75 kinds of microorganisms including Helicobacter showed weak positives, when 1 pg of DNA was input. Other microorganisms gave negative signals even when 100 pg of DNA was used for PCR. When compared with a Nested-PCR system to detect the urease A-subunit gene performed by a commercial reference laboratory, the results obtained (sensitivity 93.3% and specificity 73.3%) was almost equivalent. The PCR-MWP was rapid and easy for the detection of H. pylori DNA in gastric juice specimen.
Tamatsukuri
S
S
Nippon Roche K.K., PCR Dept.
Weiss
J
J
Hirota
K
K
Kawamata
O
O
Shiratori
Y
Y
Yoshida
H
H
Omata
M
M
D004269:DNA, Bacterial / Q000032:analysis*; D005750:Gastric Juice / Q000382:microbiology*; D016480:Helicobacter pylori / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D010957:Plasmids; D016133:Polymerase Chain Reaction / Q000379:methods*; D012336:RNA, Ribosomal, 16S; D012680:Sensitivity and Specificity
D004269:DNA, Bacterial; D012336:RNA, Ribosomal, 16S