Quantitation of Helicobacter pylori in the stomach using quantitative polymerase chain reaction assays.
BACKGROUND
The density of Helicobacter pylori is thought to correlate with the degree of inflammation and thus indirectly with the outcome of the infection. Rapid quantitative assays of H. pylori in gastric or duodenal mucosa are lacking. The aim was to develop quantitative assays using the polymerase chain reaction to assess the quantity of H. pylori in the gastric mucosa.


METHODS
Competitive PCR was based on coamplification of a segment of the ureC sequence and an internal control using a single set of primers. PCR products were quantified colorimetrically by an enzyme-linked immunosorbent assay and compared with known quantities of the internal control standard added to the PCR reaction. The highly sensitive, noncompetitive PCR assay does not use coamplification and measures the amplified DNA sequence using a flash-type luminescent tag and a specific probe. The mouse infected model using H. pylori strain SS-1 was used to develop the assays.


RESULTS
Quantification of H. pylori using either the competitive or noncompetitive PCR was reliable, highly sensitive and specific.


CONCLUSIONS
The ability to rapidly quantitate H. pylori from gastric mucosa should be useful to investigate the role of H. pylori density and infection outcome, as well as to monitor the effectiveness of antibiotic treatment or vaccines against H. pylori.
Ozpolat
B
B
Departments of Bioimmunotherapy, The University of Texas-Houston Medical School, Baylor, TX 77030, USA.
Actor
J K
JK
Rao
X M
XM
Lee
S
S
Osato
M
M
Graham
D Y
DY
Lachman
L B
LB
D000818:Animals; D002772:Cholera Toxin / Q000008:administration & dosage; D004269:DNA, Bacterial / Q000032:analysis / Q000235:genetics; D005753:Gastric Mucosa / Q000382:microbiology; D016481:Helicobacter Infections / Q000382:microbiology* / Q000517:prevention & control; D016480:Helicobacter pylori / Q000235:genetics*; D051379:Mice; D008810:Mice, Inbred C57BL; D016133:Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D013047:Specific Pathogen-Free Organisms; D013270:Stomach / Q000382:microbiology*; D014510:Urease / Q000008:administration & dosage; D014611:Vaccination
D004269:DNA, Bacterial; D002772:Cholera Toxin; D014510:Urease