Real-time polymerase chain reaction detection of Neisseria meningitidis in formalin-fixed tissues from sudden deaths

Diagn Microbiol Infect Dis. 2008 Apr;60(4):339-46. doi: 10.1016/j.diagmicrobio.2007.10.014. Epub 2008 Feb 20.

Abstract

Accurate identification of meningococcal sudden deaths is needed to avoid underestimation of the true incidence of the disease. This study analyzed the usefulness of a real-time polymerase chain reaction (PCR) protocol using MGB (3'-minor groove binder) probes to detect Neisseria meningitidis in formalin-fixed paraffin-embedded tissues from sudden deaths where a meningococcal fulminating infection was suspected. The protocol included detection of meningococcal DNA (ctrA gene), multiplex B/C PCR serogrouping (siaD gene), and rapid confirmation of PCR products by microcapillary electrophoresis. Sixty-nine tissues from 15 culture-confirmed meningococcal sudden deaths were analyzed (positive cases). Validation studies were performed. In each positive case, both the ctrA and the B/C siaD genes were detected. The ctrA was detected in 81.2% of the samples, whereas the serogroup (B or C) was identified in 44.9% of them. Therefore, this protocol may improve nonculture diagnosis and case ascertainment in meningococcal disease deaths, particularly when formalin-fixed tissues are the only available specimen.

Publication types

  • Evaluation Study

MeSH terms

  • Adolescent
  • Adult
  • Aged
  • Bacterial Proteins / genetics
  • Child
  • Child, Preschool
  • DNA Primers / genetics
  • Death, Sudden / etiology*
  • Electrophoresis, Capillary
  • Female
  • Genotype
  • Humans
  • Infant
  • Male
  • Meningococcal Infections / diagnosis*
  • Middle Aged
  • Neisseria meningitidis / genetics
  • Neisseria meningitidis / isolation & purification*
  • Polymerase Chain Reaction / methods*
  • Sensitivity and Specificity

Substances

  • Bacterial Proteins
  • DNA Primers