J Korean Med Sci.  1991 Dec;6(4):338-347. 10.3346/jkms.1991.6.4.338.

Development of epidemiological method for the Helicobacter pylori by polymerase chain reaction

• Lee WK
• Cho MJ
• Choi HJ
• Rhee KH

• KMID: 1421876
• DOI: http://doi.org/10.3346/jkms.1991.6.4.338

Abstract

The polymerase chain reaction was used to develop a method for the detection of Helicobacter pylori, a causative agent of gastritis, as well as for the elucidation of its mode of transmission. A genomic library of Helicobacter pylori DNA in Escherichia coli JM109 was constructed by cloning Hind III-digested DNA fragments into plasmid vector pUC18. The nucleotide sequences from seven recombinant clones were determined and five sets of oligonucleotide primers were synthesized on the basis of the sequences from five clones (B4, B9, B10, C15 and I22). The PCR amplifications with these primers were performed using DNA samples from five strains of Helicobacter pylori, two Campylobacter spp. and eleven species of enteric bacteria. Amplifications of the target DNA fragments in all of 5 strains of Helicobacter pylori were observed from the PCR with primers derived from clone B4, B9, C15 and I22. When the specificity was checked with the DNA samples from 13 other bacteria as template DNA for the PCR, specific amplification that produced the correct size of the target DNA of Helicobacter pylori was shown only in the PCR with primers derived from clone B9 and C15. The detection limit in the PCR amplification, determined by the heat-lysis method, was 500 cells of Helicobacter pylori.

Keyword

Helicobacter pylori; Genomic library; Nucleotide sequence; Polymerase chain reaction; Gastritis

MeSH Terms

Base Sequence
DNA, Bacterial/*analysis
DNA, Recombinant
Genomic Library
Helicobacter Infections/diagnosis
Helicobacter pylori/*genetics/isolation & purification
Humans
Molecular Sequence Data
Polymerase Chain Reaction
Sensitivity and Specificity
DNA, Bacterial
DNA, Recombinant