title: Multiplex 5' nuclease-quantitative PCR for diagnosis of relapsing fever in a large Tanzanian cohort.
abstract: Relapsing fever (RF) is caused by tick- and louse-borne Borrelia spp., is characterized by recurrent fever, and is often misdiagnosed as malaria. Because of submicroscopic bacteremia, microscopy can be insensitive between febrile bouts. We designed a multiplex quantitative PCR (qPCR) assay to distinguish RF Borrelia from Plasmodium falciparum and P. vivax. The assay specifically (100%) amplified pathogenic RF Borrelia (1 copy/reaction). We then tested blood from participants within a Tanzanian cohort assessed at scheduled intervals and with fever. Among 8,617 blood samples from 2,057 participants surveyed routinely, 7 (0.08%) samples and 7 (0.3%) participants had RF DNA (median, 4.4 × 10(3) copies/ml). Of 382 samples from 310 febrile persons, 15 (3.9%) samples from 13 (4.2%) participants had RF DNA (median, 7.9 × 10(2) copies/ml). Five (1.3%) samples from 4 (1.3%) participants were found to harbor Borrelia by microscopy. We conclude that multiplex qPCR holds promise for improved clinical diagnosis and epidemiologic assessment of RF.
authors.affiliation: Johns Hopkins University School of Medicine, 720 Rutland Avenue, Ross 624, Baltimore, MD 21205, USA. mreller1@jhmi.edu
authors.forename: Megan E
authors.initials: ME
authors.lastname: Reller
authors.forename: Emily G
authors.initials: EG
authors.lastname: Clemens
authors.forename: Steve E
authors.initials: SE
authors.lastname: Schachterle
authors.forename: George A
authors.initials: GA
authors.lastname: Mtove
authors.forename: David J
authors.initials: DJ
authors.lastname: Sullivan
authors.forename: J Stephen
authors.initials: JS
authors.lastname: Dumler
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001898:Borrelia / Q000235:genetics / Q000302:isolation & purification*; D002648:Child; D002675:Child, Preschool; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D005260:Female; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D011247:Pregnancy; D015995:Prevalence; D012061:Relapsing Fever / Q000175:diagnosis* / Q000453:epidemiology; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D013636:Tanzania / Q000453:epidemiology; D055815:Young Adult
chemical_list: D004269:DNA, Bacterial