Amplification of rDNA loci to detect and type Neisseria meningitidis and other eubacteria

Mol Cell Probes. 1993 Feb;7(1):7-17. doi: 10.1006/mcpr.1993.1002.

Abstract

In 1991-92, Neisseria meningitidis group C was isolated from the blood of eight students in Urbana, Illinois, USA, and from the cerebrospinal fluid of one student from a nearby community, Decatur, Illinois. These and other bacterial species were analysed by PCR fingerprinting using primers selected from the ribosomal (r)DNA loci. A rDNA primer pair spanning a region within the 16S rDNA amplified a predicted 280 base pair (bp) DNA fragment from Neisseria spp. and fragments of different sizes for other genera. This primer pair specifically detected a carrier of N. meningitidis in a small clinical battery. Identity of the fragment was confirmed by restriction endonuclease analysis. A 600 bp fragment was also amplified from the 16S-23S internal transcribed spacer (ITS) of N. meningitidis; amplification from six other genera yielded different-sized fragments. Digestion of the ITS fragment from N. meningitidis with Alu I revealed three patterns; pattern I was found only for serogroup C isolates, and it was the dominant pattern among recent isolates with the exception of the one from Decatur. The isolate from Decatur yielded pattern III which suggested a non-clonal relationship to the seven isolates from Urbana. Patterns II and III were more prevalent in isolates from the 1960's and 1980's. PCR-based analysis of these loci can complement the techniques which are currently used for the detection and typing of these and other eubacteria.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Adult
  • Bacterial Typing Techniques*
  • Base Sequence
  • Carrier State / diagnosis
  • Carrier State / microbiology
  • DNA, Bacterial / genetics*
  • DNA, Ribosomal / genetics*
  • Disease Outbreaks
  • Genes, Bacterial
  • Humans
  • Illinois / epidemiology
  • Meningitis, Meningococcal / epidemiology
  • Meningitis, Meningococcal / microbiology*
  • Molecular Sequence Data
  • Neisseria meningitidis / classification
  • Neisseria meningitidis / genetics
  • Neisseria meningitidis / isolation & purification*
  • Polymerase Chain Reaction*
  • RNA, Ribosomal, 16S / genetics
  • RNA, Ribosomal, 23S / genetics

Substances

  • DNA, Bacterial
  • DNA, Ribosomal
  • RNA, Ribosomal, 16S
  • RNA, Ribosomal, 23S