Detection of Helicobacter pylori in gastric biopsy tissue by polymerase chain reaction.
To evaluate the sensitivity of a polymerase chain reaction (PCR) assay using nested primers in detecting Helicobacter pylori, gastric tissue biopsy specimens were collected on endoscopy from 17 patients with a duodenal ulcer. DNA was extracted by phenol/chloroform treatment or boiling in water, and then subjected to a nested PCR using two primer pairs from the urease gene of Helicobacter pylori. Fourteen of the 17 patients were positive for Helicobacter pylori using DNA samples extracted by either method. The PCR results correlated well with the results of an enzyme immunoassay to detect IgG antibody. However, there were two culture negative patients. The three PCR negative patients were both culture negative and serologically negative. DNA from 9 of the 14 patients was randomly selected and subjected to semiquantification by serial dilutions, and then PCR. The results showed that phenol/chloroform extraction yielded 10-1000 times more DNA than the boiling method. It is concluded that the PCR assay is a rapid and sensitive method for detecting Helicobacter pylori, and that phenol/chloroform extraction is superior to simple boiling in obtaining DNA samples for PCR.
Wang
J T
JT
Department of Internal Medicine, National Taiwan University Hospital, Taipei.
Lin
J T
JT
Sheu
J C
JC
Yang
J C
JC
Chen
D S
DS
Wang
T H
TH
D000907:Antibodies, Bacterial / Q000097:blood; D001483:Base Sequence; D001706:Biopsy; D004269:DNA, Bacterial / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification*; D004381:Duodenal Ulcer / Q000150:complications / Q000382:microbiology*; D016481:Helicobacter Infections / Q000150:complications / Q000175:diagnosis / Q000382:microbiology*; D016480:Helicobacter pylori / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D006801:Humans; D007074:Immunoglobulin G / Q000097:blood; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011706:Pyloric Antrum / Q000382:microbiology*; D012680:Sensitivity and Specificity; D012689:Sequence Homology, Nucleic Acid
D000907:Antibodies, Bacterial; D004269:DNA, Bacterial; D007074:Immunoglobulin G