Direct detection of leptospiral material in human postmortem samples

Paul D. Brown, Diane G. Carrington, Claudia Gravekamp, Herman Van De Kemp, Charles N. Edwards, Stephen R. Jones, Patsy R. Prussia, Stuart Garriques, Wiepko J. Terpstra, Paul N. Levett

Research output: Contribution to journalArticle

34 Scopus citations

Abstract

Leptospiral culture, direct immunofluorescence, and the polymerase chain reaction (PCR) were used to detect leptospiral material in postmortem specimens collected from eight patients who died of leptospirosis. Diagnosis of leptospiral infection was based on clinical summary (premortem) and confirmed by serological analysis and/or culture of leptospires. Leptospiral culture was the least sensitive technique, yielding two isolates (3%) from 65 samples. Both isolates were from the aqueous humour and cerebrospinal fluid of the same patient. Direct immunofluorescence was of intermediate sensitivity for detection of leptospires, confirming the presence of leptospires in 11% (2 of 18) of tissue samples from three patients. PCR analysis was the most sensitive technique for detection of leptospiral material in tissue samples, being positive in 20% (11 of 56) of samples from eight patients. Both samples (cerebellum and liver) positive by immunofluorescence were also positive by PCR. The sensitivity of the PCR assay was 1-10 leptospires ml-1 sample, and the assay was specific for Leptospira pathogenic species. Multi-system involvement was indicated based on successful amplification of leptospiral DNA from more than one tissue sample, which corroborated with the clinical and pathologic findings. The results suggest that in acute and/or fatal leptospirosis, the pathogenesis of the pathologic features are related to the presence of the organisms in the tissues. In conclusion, PCR combined with serology appears to be a useful tool for diagnosis of leptospirosis and may be invaluable in epidemiological studies.

Original languageEnglish (US)
Pages (from-to)581-586
Number of pages6
JournalResearch in Microbiology
Volume154
Issue number8
DOIs
StatePublished - Oct 2003
Externally publishedYes

Keywords

  • Detection
  • Immunofluorescence
  • Leptospira
  • PCR

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology

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