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KMID : 0357319940290010041
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Journal of the Korean Society for Microbiology
1994 Volume.29 No. 1 p.41 ~ p.56
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Diagnosis of Helicobacter pylori Infection by Polymerase Chain Reaction Using the Gene of 120KD Protein of Helicobacter pylori as Target Gene
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이광호
류항희/조명제/전영석/이우곤/김경희/백승철/강경희/초휴진
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Abstract
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Helicobacter pylori is the etiologic agent of type B chronic gastritis and strongly associated with peptic ulcer as well as gastric cancer. Accurate and reliable methods to detect H. pylori is necessary for research and clinical diagnosis. The
polymerase chain reaction(PCR) was designed to amplify Helicobcter pylori gene encoding 120kd protein which is a serologically specific component of Helicobacter pylori. Five primers sets were schemed to cover 3'-region of the gene encoding 120kd
protein and to amplifiy DNA fragments of from 300 to 400 nucleotides in size. Of 5 primers sets, Hel-2 primers set was confirmed to amplify the Helicobacter pylori gene sensitively and specifically when tested by 5 Helicobacter pylori isoiates
and
13
enteric bacteria. PCR using Hel-2 primers set could detect as little as 1pg DNA and 10 colony forming units of Helicobacter pylori. All of 60 specimens which were Helicobacter pylori-culture positive and 21(26.6%) of 79 specimens which were
culture
and
urease test negative showed the specifically amplified DNA fragment at the PCR , indicating that the PCR can detect a small number of organism on gastric biopsy samples. PCR assay also detected 1*10E4 colony forming units of Helicobacter pylori
in
stool
suspension treated with DE52 gel in order to remove the inhibitory substance of PCR in stool.
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