TY - JOUR

T1 - 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

AU - Petersen, R.F.

AU - Harrington, Clare Sarah

AU - Kortegaard, Hanne Ellen

AU - On, S.L.W.

PY - 2007

Y1 - 2007

N2 - 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 profilegroups 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 ampylobacters. 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.

AB - 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 profilegroups 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 ampylobacters. 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.

KW - Former LIFE faculty

KW - 16s PCR-DGGE

KW - culture-independent identification

KW - Epsilobacteria

KW - saliva samples

U2 - 10.1111/j.1365-2672.2007.03515.x

DO - 10.1111/j.1365-2672.2007.03515.x

M3 - Journal article

VL - 103

SP - 2601

EP - 2615

JO - Proceedings of the Society for Applied Bacteriology

JF - Proceedings of the Society for Applied Bacteriology

SN - 0370-1778

IS - 6

ER -