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A PCR‐DGGE method for detection and identification of Campylobacter , Helicobacter , Arcobacter and related Epsilobacteria and its application to saliva samples from humans and domestic pets
Author(s) -
Petersen R.F.,
Harrington C.S.,
Kortegaard H.E.,
On S.L.W.
Publication year - 2007
Publication title -
journal of applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.889
H-Index - 156
eISSN - 1365-2672
pISSN - 1364-5072
DOI - 10.1111/j.1365-2672.2007.03515.x
Subject(s) - arcobacter , campylobacter , saliva , microbiology and biotechnology , biology , helicobacter , campylobacter jejuni , identification (biology) , feces , bacteria , helicobacter pylori , genetics , biochemistry , botany
Aims: To develop a PCR‐denaturing gradient gel electrophoresis (PCR‐DGGE) method for the detection and identification of Campylobacter , Helicobacter and Arcobacter species (Epsilobacteria) in clinical samples and evaluate its efficacy on saliva samples from humans and domestic pets. Methods and Results: A semi‐nested PCR was developed to allow sensitive detection of all Epsilobacteria, with species separation undertaken by DGGE. A database was constructed in BioNumerics using 145 strains covering 51 Campylobacter , Arcobacter and Helicobacter taxa; Nineteen distinct DGGE profile‐groups were distinguished. This approach detected Epsilobacteria in all saliva samples collected from humans, cats and dogs, and identified Campylobacter concisus and/or Campylobacter gracilis in the human samples. The pet animal samples were taken from individuals with oral/dental diseases; PCR‐DGGE identified up to four different species in each sample. The most common species detected included Wolinella succinogenes , Arcobacter butzleri and two hitherto uncultured campylobacters. The enteropathogen Campylobacter lari was also found. Conclusions: PCR combined with DGGE is a useful tool for direct detection and preliminary identification of Epsilobacteria in the oral cavity of humans and small animals. Significance and Impact of the Study: The PCR‐DGGE method should allow determination of the true prevalence and diversity of Epsilobacteria in clinical and other samples. Contact with the oral cavity of domestic pets may represent a route of transmission for epsilobacterial enteric diseases.