Amplification of rDNA loci to detect and type Neisseria meningitidis and other eubacteria.
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.
McLaughlin
G L
GL
Department of Veterinary Pathobiology, School of Veterinary Medicine, Purdue University, West Lafayette, IN 47907.
Howe
D K
DK
Biggs
D R
DR
Smith
A R
AR
Ludwinski
P
P
Fox
B C
BC
Tripathy
D N
DN
Frasch
C E
CE
Wenger
J D
JD
Carey
R B
RB
D000328:Adult; D015373:Bacterial Typing Techniques; D001483:Base Sequence; D002353:Carrier State / Q000175:diagnosis / Q000382:microbiology; D004269:DNA, Bacterial / Q000235:genetics*; D004275:DNA, Ribosomal / Q000235:genetics*; D004196:Disease Outbreaks; D005798:Genes, Bacterial; D006801:Humans; D007087:Illinois / Q000453:epidemiology; D008585:Meningitis, Meningococcal / Q000453:epidemiology / Q000382:microbiology*; D008969:Molecular Sequence Data; D009345:Neisseria meningitidis / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D012338:RNA, Ribosomal, 23S / Q000235:genetics
D004269:DNA, Bacterial; D004275:DNA, Ribosomal; D012336:RNA, Ribosomal, 16S; D012338:RNA, Ribosomal, 23S