Detection of
            <i>Helicobacter</i>
            Colonization of the Murine Lower Bowel by Genus-Specific PCR-Denaturing Gradient Gel Electrophoresis

Grehan, Martin; Tamotia, Gauri; Robertson, Bronwyn R.; Mitchell, Hazel M.

doi:10.1128/aem.68.10.5164-5166.2002

Cited by 16 publications

(20 citation statements)

References 20 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Using fecal PCR-denaturing gradient gel electrophoresis, Grehan et al surveyed eight mice from three suppliers in Australia (14). They reported that four of eight mice were colonized with H. ganmani, two of eight had H. bilis, three of eight had H. hepaticus, and one of eight had two unique isolates, in addition to H. ganmani.…”

Section: Discussionmentioning

confidence: 99%

Enterohepatic Helicobacter Species Are Prevalent in Mice from Commercial and Academic Institutions in Asia, Europe, and North America

Taylor

Nambiar

et al. 2007

J Clin Microbiol

View full text Add to dashboard Cite

The discovery of Helicobacter hepaticus and its role in hepatitis, hepatocellular carcinoma, typhlocolitis, and lower-bowel carcinoma in murine colonies was followed by the isolation and characterization of other Helicobacter spp. involved in enterohepatic disease. Colonization of mouse colonies with members of the family Helicobacteriaceae has become an increasing concern for the research community.

show abstract

Section: Discussionmentioning

confidence: 99%

Enterohepatic Helicobacter Species Are Prevalent in Mice from Commercial and Academic Institutions in Asia, Europe, and North America

Taylor

Nambiar

et al. 2007

J Clin Microbiol

View full text Add to dashboard Cite

show abstract

“…PCRrestriction fragment length polymorphism analysis is efficient when it is used with genomic DNA from cultured organisms, but its value is uncertain for the direct detection of Helicobacter in animal tissues, in which colonization with more than one Helicobacter spp. can occur (13,27). Consequently, cultureindependent methods for the direct detection and identification of species of this genus in biological samples are important.…”

Section: Discussionmentioning

confidence: 99%

High Prevalence of Helicobacter Species Detected in Laboratory Mouse Strains by Multiplex PCR-Denaturing Gradient Gel Electrophoresis and Pyrosequencing

et al. 2004

View full text Add to dashboard Cite

Rodent models have been developed to study the pathogenesis of diseases caused by Helicobacter pylori, as well as by other gastric and intestinal Helicobacter spp., but some murine enteric Helicobacter spp. cause hepatobiliary and intestinal tract diseases in specific inbred strains of laboratory mice. To identify these murine Helicobacter spp., we developed an assay based on PCR-denaturing gradient gel electrophoresis and pyrosequencing. Nine strains of mice, maintained in four conventional laboratory animal houses, were assessed for Helicobacter sp. carriage. Tissue samples from the liver, stomach, and small intestine, as well as feces and blood, were collected; and all specimens (n ‫؍‬ 210) were screened by a Helicobacter genus-specific PCR. Positive samples were identified to the species level by multiplex denaturing gradient gel electrophoresis, pyrosequencing, and a H. ganmani-specific PCR assay. Histologic examination of 30 tissue samples from 18 animals was performed. All mice of eight of the nine strains tested were Helicobacter genus positive; H. bilis, H. hepaticus, H. typhlonius, H. ganmani, H. rodentium, and a Helicobacter sp. flexispira-like organism were identified. Helicobacter DNA was common in fecal (86%) and gastric tissue (55%) specimens, whereas samples of liver tissue (21%), small intestine tissue (17%), and blood (14%) were less commonly positive. Several mouse strains were colonized with more than one Helicobacter spp. Most tissue specimens analyzed showed no signs of inflammation; however, in one strain of mice, hepatitis was diagnosed in livers positive for H. hepaticus, and in another strain, gastric colonization by H. typhlonius was associated with gastritis. The diagnostic setup developed was efficient at identifying most murine Helicobacter spp.

show abstract

“…Thermal cycling conditions were 94°C for 5 min and 35 cycles of 94°C for 5 s, 57°C for 5 s, and 72°C for 30 s. Twenty-microliter PCR products were examined on a 1.5% agarose gel. PCR-denaturing gradient gel electrophoresis (DGGE) analysis was conducted according to a method described previously by Grehan et al (11). Individual DGGE bands were excised and sequenced.…”

mentioning

confidence: 99%

Nongastric Helicobacter Species Detected in the Intestinal Tract of Children

et al. 2006

Self Cite

View full text Add to dashboard Cite

The presence of members of the family Helicobacteraceae in the colon of children was investigated using PCR, denaturing gradient gel electrophoresis, and fluorescent in situ hybridization. The rate of detection of species belonging to the Helicobacteraceae family in children with inflammatory bowel disease (92%) and irritable bowel syndrome (100%) was found to be significantly higher than that in healthy children (25%) (P < 0.05).Helicobacter species are gram-negative curved to spiralshaped bacteria that naturally colonize the mucus layer of the gastrointestinal tract of a range of mammalian hosts (15). Based on their preferential site of colonization, species of the Helicobacter genus have been divided into gastric and enterohepatic subgroups. Helicobacters in the gastric group, represented by Helicobacter pylori, have been associated with gastritis and other gastric disorders (15, 16). Enterohepatic Helicobacter species, which colonize the intestine and hepatobiliary system, have been linked to chronic hepatic and intestinal diseases (9, 16) as well as inflammatory bowel disease (IBD) in immunodeficient rodents (10, 13). The role of Helicobacter species in human IBD is currently controversial (4,5,19).In this study, we assessed the presence and spatial distribution of members of the Helicobacteraceae family (genera Helicobacter and Wolinella) in the colon of 21 children (16/21 male, aged 3 to 15 years) undergoing diagnostic colonoscopy and determined their association with colonic inflammation. The H. pylori status of all children, as assessed by histological examination and rapid urease testing of gastric biopsies, showed that 20/21 children were negative for H. pylori.Three colonic biopsies were collected from each child at colonoscopy. One biopsy was fixed in 10% neutral formalin for routine histological examination, the second biopsy was fixed in Carnoy's fixative for fluorescent rRNA in situ hybridization (FISH) analysis and Alcian blue-periodic acid-Schiff (PAS) staining, and the third biopsy was placed directly into 600 l of lysis buffer (Gentra Systems, Minneapolis, Minn.) for DNA extraction.DNA was extracted from freshly collected biopsy tissues using the PUREGENE DNA extraction kit (Gentra Systems, Minneapolis, Minn.). The16S rRNA gene of members of the family Helicobacteraceae was amplified by PCR using primers H276f and H676r with modified thermal cycle conditions (18). Hot-start PCRs were performed in a 25-l volume containing 10 pmol of each primer, 1ϫ PCR buffer (Fisher Biotech), 200 nM concentrations of each nucleotide triphosphate, 1.5 mM MgCl 2 , and 200 to 250 ng of extracted DNA. Thermal cycling conditions were 94°C for 5 min and 35 cycles of 94°C for 5 s, 57°C for 5 s, and 72°C for 30 s. Twenty-microliter PCR products were examined on a 1.5% agarose gel. PCR-denaturing gradient gel electrophoresis (DGGE) analysis was conducted according to a method described previously by Grehan et al. (11). Individual DGGE bands were excised and sequenced.Slides prepared from biopsies fixed in Carnoy's fi...

show abstract

Detection of Helicobacter Colonization of the Murine Lower Bowel by Genus-Specific PCR-Denaturing Gradient Gel Electrophoresis

Cited by 16 publications

References 20 publications

Enterohepatic Helicobacter Species Are Prevalent in Mice from Commercial and Academic Institutions in Asia, Europe, and North America

Enterohepatic Helicobacter Species Are Prevalent in Mice from Commercial and Academic Institutions in Asia, Europe, and North America

High Prevalence of Helicobacter Species Detected in Laboratory Mouse Strains by Multiplex PCR-Denaturing Gradient Gel Electrophoresis and Pyrosequencing

Nongastric Helicobacter Species Detected in the Intestinal Tract of Children

Contact Info

Product

Resources

About