title: Multiplex real-time PCR diagnostic of relapsing fevers in Africa.
abstract: BACKGROUND
  In Africa, relapsing fever borreliae are neglected arthropod-borne pathogens causing mild to deadly septicemia and miscarriage. The closely related Borrelia crocidurae, Borrelia duttonii, Borrelia recurrentis and Borrelia hispanica are rarely diagnosed at the species level, hampering refined epidemiological and clinical knowledge of the relapsing fevers. It would be hugely beneficial to have simultaneous detection and identification of Borrelia to species level directly from clinical samples.
  
  
  RESULTS
  We designed a multiplex real-time PCR protocol targeting the 16S rRNA gene detecting all four Borrelia, the glpQ gene specifically detecting B. crocidurae, the recN gene specifically detecting B. duttonii/B. recurrentis and the recC gene specifically detecting B. hispanica. Compared to combined 16S rRNA gene and flaB gene sequencing as the gold standard, multiplex real-time PCR analyses of 171 Borrelia-positive and 101 Borrelia-negative control blood specimens yielded 100% sensitivity and specificity for B. duttonii/B. recurrentis and B. hispanica and 99% sensitivity and specificity for B. crocidurae.
  
  
  CONCLUSIONS
  The multiplex real-time PCR developed in this study is a rapid technique for both molecular detection and speciation of relapsing fever borreliae from blood in Africa. It could be incorporated in point-of-care laboratory to confirm diagnosis and provide evidence of the burden of infection attributed to different species of known or potentially novel relapsing fever borreliae.
authors: Aix Marseille Université, URMITE, UMR63, CNRS 7278, IRD 198, Inserm 1095, Marseille, France.
authors: Haitham
authors: H
authors: Elbir
authors: Mireille
authors: M
authors: Henry
authors: Georges
authors: G
authors: Diatta
authors: Oleg
authors: O
authors: Mediannikov
authors: Cheikh
authors: C
authors: Sokhna
authors: Adama
authors: A
authors: Tall
authors: Cristina
authors: C
authors: Socolovschi
authors: Sally J
authors: SJ
authors: Cutler
authors: Kassahum D
authors: KD
authors: Bilcha
authors: Jemal
authors: J
authors: Ali
authors: Dayana
authors: D
authors: Campelo
authors: Steven C
authors: SC
authors: Barker
authors: Didier
authors: D
authors: Raoult
authors: Michel
authors: M
authors: Drancourt
mesh_terms: D000349:Africa; D001431:Bacteriological Techniques / Q000379:methods; D001769:Blood / Q000382:microbiology; D001898:Borrelia / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012061:Relapsing Fever / Q000175:diagnosis*; D012680:Sensitivity and Specificity; D013997:Time Factors
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