Multiplex 5' nuclease-quantitative PCR for diagnosis of relapsing fever in a large Tanzanian cohort - PubMed (original) (raw)

Multiplex 5' nuclease-quantitative PCR for diagnosis of relapsing fever in a large Tanzanian cohort

Megan E Reller et al. J Clin Microbiol. 2011 Sep.

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.

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Figures

Fig. 1.

Fig. 1.

Quantitation of relapsing fever Borrelia by quantitative PCR among participants evaluated with febrile illness versus routine surveillance.

Fig. 2.

Fig. 2.

Alignment and phylogenetic analysis of flaB amplified from the blood of 7 Tanzanian patients with relapsing fever caused by Borrelia, represented by RF4, RF6, RF8, RF16, RF19, RF20, and RF23, with GenBank accession numbers in parentheses. Borrelia miyamotoi is included to represent the outgroup. The data represent the numbers of bootstrap replicates out of 1,000 iterations for each branch. The inset scale represents the number of base pair substitutions per 1,000 nucleotides.

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