Lab Anim Res.  2015 Jun;31(2):86-92. 10.5625/lar.2015.31.2.86.

Comparison of three diagnostic assays for the identification of Helicobacter spp. in laboratory dogs

• Hong S
• Chung Y
• Kang WG
• Choi YS
• Kim O

• KMID: 2053682
• DOI: http://doi.org/10.5625/lar.2015.31.2.86

Abstract

A number of Helicobacter species may confound experimental data because of their association with disease progressing in various kinds of laboratory animals. Screening of Helicobacter species is particularly desirable, because they are prevalent in commercial and research animal facilities. The aim of the present study was to compare three diagnostic methods [e.g. Helicobacter stool antigen kit (HpSA), polymerase chain reaction (PCR) and rapid urease test (RUT)] for the identification of Helicobacter spp. in stools or gastric biopsy specimens collected from eight dogs suffering from gastritis. The gastroscopic biopsy specimens were tested using RUT and PCR, while stool specimens were evaluated using both HpSA and PCR. DNAs from the gastric biopsies and stool specimens were analyzed by both a consensus PCR that amplified the RNA polymerase beta-subunit-coding gene (rpoB) of Helicobacter spp. and a species-specific PCR to amplify the urease B gene of Helicobacter heilmannii, Helicobacter pylori, and Helicobacter felis. Helicobacter spp. were detected in 62.5% of the dogs, while H. heilmannii and H. felis were identified in 37.5 and 25% of the dogs, respectively. The HpSA did not efficiently detect Helicobacter spp. in the stool samples compared to the RUT and PCR assays, both of which successfully detected Helicobacter spp. in the two sample types. Finally, we recommend that consensus PCR with stool specimens could be used before the species-specific PCR for identifying Helicobacter species in laboratory dogs.

Keyword

Helicobacter; stool antigen kit; stool; PCR; dog

MeSH Terms

Animals
Animals, Laboratory
Biopsy
Cats
Consensus
DNA
DNA-Directed RNA Polymerases
Dogs*
Felis
Gastritis
Helicobacter felis
Helicobacter heilmannii
Helicobacter pylori
Helicobacter*
Mass Screening
Polymerase Chain Reaction
Urease
DNA
DNA-Directed RNA Polymerases
Urease

Figure

• 

  Figure 1 Amplification of Helicobacter rpoB DNAs from consensus PCR with gastroscopic biopsy specimens was identified on a 1.2% agarose gel electrophoresis. M: Size marker, P: Positive control (H. pylori), N: Distilled water, Lane 1~8: Gastroscopic biopsy specimen of dog No. 1~8.

• 

  Figure 2 Results of the multiplex species-specific PCR to amplify urease B gene for H. heilmannii, H. pylori and H. felis. M: Size marker, Ph: Positive control (H. heilmannii), Pp: Positive control (H. pylori), Pf: Positive control (H. felis), N: Distilled water, Lane 1~8: Gastroscopic biopsy specimen of dog No. 1~8.

• 

  Figure 3 Scheme of detection of Helicobacter species in the canine stools.

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