1. Amplification of rDNA loci to detect and type Neisseria meningitidis and other eubacteria.
- Author
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McLaughlin GL, Howe DK, Biggs DR, Smith AR, Ludwinski P, Fox BC, Tripathy DN, Frasch CE, Wenger JD, and Carey RB
- Subjects
- Adult, Base Sequence, Carrier State diagnosis, Carrier State microbiology, Disease Outbreaks, Genes, Bacterial, Humans, Illinois epidemiology, Meningitis, Meningococcal epidemiology, Molecular Sequence Data, Neisseria meningitidis classification, Neisseria meningitidis genetics, RNA, Ribosomal, 16S genetics, RNA, Ribosomal, 23S genetics, Bacterial Typing Techniques, DNA, Bacterial genetics, DNA, Ribosomal genetics, Meningitis, Meningococcal microbiology, Neisseria meningitidis isolation & purification, Polymerase Chain Reaction
- 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.
- Published
- 1993
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