Comparison of two PCR methods for detection of Leptospira interrogans in formalin-fixed and paraffin-embedded tissues

Aleco D'Andrea, Yarelys Zamora Martinez, Rosa Alduina, Vincenzo Monteverde, Carmen Fernandez Molina, Maria Vitale

Research output: Contribution to journalArticlepeer-review

Abstract

In this study we compared two polymerase chain reaction (PCR) methods using either 16S ribosomal RNA (rRNA) or 23S rRNA gene primers for the detection of different Leptospira interrogans serovars. The performance of these two methods was assessed using DNA extracted from bovine tissues previously inoculated with several bacterial suspensions. PCR was performed on the same tissues before and after the formalin-fixed, paraffin-embedding procedure (FFPE tissues). The 23S rDNA PCR detected all fresh and FFPE positive tissues while the 16S rDNA-based protocol detected primarily the positive fresh tissues. Both methods are specific for pathogenic L. interrogans. The 23S-based PCR method successfully detected Leptospira in four dubious cases of human leptospirosis from archival tissue specimens and one leptospirosis-positive canine specimen. A sensitive method for leptospirosis identification in FFPE tissues would be a useful tool to screen histological specimen archives and gain a better assessment of human leptospirosis prevalence, especially in tropical countries, where large outbreaks can occur following the rainy season.

Original languageEnglish
Pages (from-to)85-88
Number of pages4
JournalMemorias do Instituto Oswaldo Cruz
Volume107
Issue number1
Publication statusPublished - Feb 2012

Keywords

  • Diagnostics
  • FFPE tissue
  • Leptospira
  • PCR

ASJC Scopus subject areas

  • Microbiology (medical)

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