title: A novel Campylobacter jejuni sequence type from a culture-negative patient in the Gambia.
abstract: The introduction of molecular diagnostic methods is crucial for improved understanding of the aetiology and epidemiology of bacterial infections in communities in resource poor settings. A blood sample from a 7 month old patient diagnosed with malaria in 2001 in a Gambian outpatient clinic was reported as culture negative after it was subjected to traditional bacterial culture protocols. We re-addressed the analysis of the blood sample from this case more recently (after 6.5 years in archival storage) in pilot work establishing 16S rRNA PCR in our molecular laboratory. Initial 16S rRNA PCR results confirmed the presence of bacterial DNA in the sample. 16S rRNA sequence analysis identified the organism as Campylobacter spp. In light of the molecular evidence we successfully grew the organism using appropriate culture conditions and subsequently biochemically confirmed that the isolate was Campylobacter jejuni. PCR and DNA sequencing of a set of seven C. jejuni housekeeping genes and in silico Multilocus Sequence Typing (MLST) analysis revealed that the isolate exhibits a novel sequence type (ST) of C. jejuni (ST 2928) and belongs to ST-443 clonal complex. This study demonstrates the potential for molecular tools to enhance the diagnosis of bacterial infections, which remain a major killer globally, not least in children in the developing world. Improvements in diagnostics are needed, and will be important not only for sick individuals but also for populations, where better measures of disease burden will contribute significantly to the improvement of public health policy.
authors.affiliation: Bacterial Diseases Programme, Medical Research Council Laboratories, Fajara, The Gambia. gmorris@mrc.gm
authors.forename: Gerard A J
authors.initials: GA
authors.lastname: Morris
authors.forename: Usman N
authors.initials: UN
authors.lastname: Ikumapayi
authors.forename: Martin
authors.initials: M
authors.lastname: Antonio
authors.forename: Stephen R C
authors.initials: SR
authors.lastname: Howie
authors.forename: Richard A
authors.initials: RA
authors.lastname: Adegbola
mesh_terms: D001483:Base Sequence; D016123:Campylobacter jejuni / Q000235:genetics / Q000302:isolation & purification*; D017931:DNA Primers; D004587:Electrophoresis, Agar Gel; D005714:Gambia; D005798:Genes, Bacterial; D006801:Humans; D016133:Polymerase Chain Reaction; D012336:RNA, Ribosomal, 16S / Q000235:genetics
chemical_list: D017931:DNA Primers; D012336:RNA, Ribosomal, 16S