Development of a genus-specific PCR assay for the molecular detection, confirmation and identification of Fusobacterium spp

Y. NaganoNorthern Ireland Public Health Laboratory, Department of Bacteriology, Belfast, Northern Ireland, UK

M. WatabeNorthern Ireland Public Health Laboratory, Department of Bacteriology, Belfast, Northern Ireland, UK

K.G. PorterDepartment of Gastroenterology, Belfast City Hospital, Belfast, Northern Ireland, UK

W.A. CoulterSchool of Dentistry, Queen’s University, The Royal Group of Hospitals, Belfast City Hospital,Belfast, Northern Ireland, UK

B.C. MillarNorthern Ireland Public Health Laboratory, Department of Bacteriology, Belfast, Northern Ireland, UK

J.S. ElbornNorthern Ireland Regional Adult Cystic Fibrosis Centre, Belfast City Hospital,Belfast, Northern Ireland, UK;Department of Respiratory Medicine, Queen’s University, Belfast City Hospital,Belfast, Northern Ireland, UK

C.E. GoldsmithNorthern Ireland Public Health Laboratory, Department of Bacteriology, Belfast, Northern Ireland, UK

P.J. RooneyNorthern Ireland Public Health Laboratory, Department of Bacteriology, Belfast, Northern Ireland, UK

A. LoughreyNorthern Ireland Public Health Laboratory, Department of Bacteriology, Belfast, Northern Ireland, UKCorrespondence[email protected]

J.E. MooreNorthern Ireland Public Health Laboratory, Department of Bacteriology, Belfast, Northern Ireland, UK

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Abstract

A genus-specific polymerase chain reaction (PCR)-based assay is developed for the detection and identification of clinically relevant Fusobacterium species, including F. nucleatum and F. necrophorum. Two 16S ribosomal DNA (rDNA) primers, FUSO1 (forward primer: 5’-GAG AGA GCT TTG CGT CC-3’ [17-mer]) and FUSO 2 (reverse primer: 5’-TGG GCG CTG AGG TTC GAC -3’ [18-mer]) are designed to target conserved regions of the 16S rDNA gene for Fusobacterium spp. Subsequent proof-of-principle studies employing this assay detected Fusobacterium spp. in the faeces of eight (10%) out of 80 patients with suspected gastrointestinal infection. This assay may be used for the genus-specific detection of Fusobacterium spp. from clinical specimens and for subsequent species identification.

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Development of a genus-specific PCR assay for the molecular detection, confirmation and identification of Fusobacterium spp

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Development of a genus-specific PCR assay for the molecular detection, confirmation and identification of Fusobacterium spp

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