Molecular detection and speciation of pathogenic Leptospira spp. in blood from patients with culture-negative leptospirosis (biomedcentral)
Background: Pathogenic Leptospira spp. present in the blood of patients with
leptospirosis during the first week of symptoms can be detected using culture
or PCR. A proportion of patients who are positive by PCR are negative by
culture. Leptospira spp. are fastidious bacteria, and we hypothesized that a
false-negative culture result may represent infection with a distinct
bacterial subset that fail to grow in standard culture medium. Methods: We
evaluated our hypothesis during a prospective study of 418 consecutive
patients presenting to a hospital in northeast Thailand with an acute febrile
illness. Admission blood samples were taken for Leptospira culture and PCR. A
single tube nested PCR that amplified a region of the rrs gene was developed
and applied, amplicons sequenced and a phylogenetic tree reconstructed.
Results: 39/418 (9%) patients were culture-positive for Leptospira spp., and
81/418 (19%) patients were culture-negative but rrs PCR-positive. The species
associated with culture-positive leptospirosis (37 L. interrogans and 2 L.
borgpetersenii) were comparable to those associated with culture-negative,
PCR-positive leptospirosis (76 L. interrogans, 4 L. borgpetersenii, 1
unidentified, possibly new species). Conclusion: Molecular speciation failed
to identify a unique bacterial subset in patients with culture-negative, PCR-
positive leptospirosis. The rate of false-negative …
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