Development of new PCR primers by comparative genomics for the detection of Helicobacter suis in gastric biopsy specimens

Helicobacter. 2014 Aug;19(4):260-71. doi: 10.1111/hel.12127. Epub 2014 Mar 28.

Abstract

Background: Although the infection rate of Helicobacter suis is significantly lower than that of Helicobacter pylori, the H. suis infection is associated with a high rate of gastric mucosa-associated lymphoid tissue (MALT) lymphoma. In addition, in vitro cultivation of H. suis remains difficult, and some H. suis-infected patients show negative results on the urea breath test (UBT).

Materials and methods: Female C57BL/6J mice were orally inoculated with mouse gastric mucosal homogenates containing H. suis strains TKY or SNTW101 isolated from a cynomolgus monkey or a patient suffering from nodular gastritis, respectively. The high-purity chromosomal DNA samples of H. suis strains TKY and SNTW101 were prepared from the infected mouse gastric mucosa. The SOLiD sequencing of two H. suis genomes enabled comparative genomics of 20 Helicobacter and 11 Campylobacter strains for the identification of the H. suis-specific nucleotide sequences.

Results: Oral inoculation with mouse gastric mucosal homogenates containing H. suis strains TKY and SNTW101 induced gastric MALT lymphoma and the formation of gastric lymphoid follicles, respectively, in C57BL/6J mice. Two conserved nucleotide sequences among six H. suis strains were identified and were used to design diagnostic PCR primers for the detection of H. suis.

Conclusions: There was a strong association between the H. suis infection and gastric diseases in the C57BL/6 mouse model. PCR diagnosis using an H. suis-specific primer pair is a valuable method for detecting H. suis in gastric biopsy specimens.

Keywords: Helicobacter suis; PCR diagnosis; comparative genomics; gastric biopsy specimen; mouse infection model; next-generation sequencing.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Biopsy
  • DNA Primers / genetics*
  • DNA, Bacterial / chemistry
  • DNA, Bacterial / genetics
  • Disease Models, Animal
  • Female
  • Gastric Mucosa / microbiology*
  • Genome, Bacterial
  • Helicobacter Infections / diagnosis*
  • Helicobacter Infections / microbiology
  • Helicobacter heilmannii / genetics
  • Helicobacter heilmannii / isolation & purification*
  • Humans
  • Macaca fascicularis
  • Mice, Inbred C57BL
  • Molecular Sequence Data
  • Pathology, Molecular / methods*
  • Polymerase Chain Reaction / methods*
  • Sequence Analysis, DNA

Substances

  • DNA Primers
  • DNA, Bacterial

Associated data

  • GENBANK/AB849014
  • GENBANK/AB849015
  • GENBANK/AB849016
  • GENBANK/AB849017
  • GENBANK/AB849018
  • GENBANK/AB849019
  • GENBANK/AB849020
  • GENBANK/AB849021