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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 culture was high, and we speculate that antibiotic pre-treatment is the most likely explanation for this.

Molecular detection and speciation of pathogenic Leptospiraspp. in blood from patients with culture-negative leptospirosis