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

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.

Detection of Helicobacter spp. in the saliva of dogs with gastritis

“…This methodology renders the PCR more sensitive and theoretically allows the identification of up to two xenogenic cells in the analysed material. At the same time the use of two pairs of primers ensures the high specificity of this method and makes it possible to reject false positive results (Singh et al 2008, Hong et al 2015. The selection of primers and PCR reaction schemes are presented in Table 1.…”

Detection of gastric Helicobacter spp. in stool samples of dogs with gastritis

“…In this study, using nested-PCR to assess stool samples, Helicobacter microorganisms were identified in 23% of animals. Hong et al (2015) obtained a higher percentage of gastric Helicobacter spp. DNA in faeces, amounting to 62.5%.…”

“…The large discrepancy in results in human and veterinary medicine may result from the small amount of gastric Helicobacter spp. in faeces, a degradation of bacterial DNA in the large intestine, the presence of polymerase inhibitors such as complex polysaccharides, and may depend on the DNA extraction method and type of PCR used (Kabir 2001, Shinozaki et al 2002, Falsafi et al 2009, Smith et al 2012, Hong et al 2015.…”

“…Helicobacter bacteria may colonize the gastrointestinal mucosa of humans, domestic animals (such as the dog, cat, pig, ferret and poultry) and wild animals (such as the cheetah or monkey) (Haesebrouck et al 2009, Casagrande Proietti et al 2010, Abdi et al 2014, Hong et al 2015. In human medicine, Helicobacter pylori has been confirmed as contributing to the development of chronic inflammation of the gastric mucosa as well as gastric and duodenal ulcers.…”

“…However, they also have a number of drawbacks. These include patient discomfort associated with carrying out a gastroscopy, obtaining biopsy samples and general anaesthesia associated with these procedures (Shinozaki et al 2002, Kubiak 2006, Haesebrouck et al 2009, Hong et al 2015.…”