title: [A method for determination of Neisseria meningitidis serogroup A, B, C and W by real-time PCR].
abstract: OBJECTIVE
  Development and testing of a real-time PCR method for detection of Neisseria meningitiis serogroup A, B, C and W DNA.
  
  
  METHODS
  Reference strains and 187 samples of cerebrospinal fluid (CSF) from meningocci meningitis patients were used in the study. Multiplex PCR was carried out in an instrument with 5 channels of fluorescent detection:
  
  
  RESULTS
  Analysis of specific serogroup loci of the genome and design of oligonucleotides for the detection of DNA of all the capsule meningococci and 4 serogroups in particular was carried out. PCR conditions were optimized; specificity was shown and analytical sensitivity was evaluated using reference strains. DNA of the following serogroups was detected during study of clinical CSF samples: A--in 103 samples (55%), B--in 45 (24%), C--in 30 (16%), W--in 5 (3%). Only DNA of meningococci capstle gene ctrA was found in 4 samples; presumably, they contained DNA of other serogroups. Multilocus sequence-typing and detection of antigenic determinants of PorA and FetA genes for 27 DNA samples of group A menincococci as well as DNA of 5 group W meningococci and 4 ungroupable was carried out.
  
  
  CONCLUSIONS
  The method proposed allows to carry out serogrouping of no less than 95% of strains or DNA samples isolated from CSF of meningococci infection patients. Combined with other recommended non-cultural methods of genotyping, it may be useful for complex characteristics of pathogenic meningococci.
authors: K O
authors: KO
authors: Mironov
authors: A E
authors: AE
authors: Platonov
authors: O P
authors: OP
authors: Dribnokhodova
authors: V I
authors: VI
authors: Kuseva
authors: G A
authors: GA
authors: Shipulin
mesh_terms: D016667:Bacterial Capsules / Q000235:genetics*; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics*; D015870:Gene Expression; D056426:Genetic Loci; D016680:Genome, Bacterial; D005838:Genotype; D006801:Humans; D008585:Meningitis, Meningococcal / Q000134:cerebrospinal fluid / Q000382:microbiology; D058885:Multilocus Sequence Typing; D060885:Multiplex Polymerase Chain Reaction; D009345:Neisseria meningitidis / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification; D018272:Porins / Q000235:genetics; D065288:Serogroup; D012703:Serotyping
chemical_list: D001425:Bacterial Outer Membrane Proteins; D004269:DNA, Bacterial; C093120:FrpB protein, bacteria; D018272:Porins; C057699:porin protein, Neisseria