Article: [A method for determination of Neisseria meningitidis serogroup A, B, C and W by real-time PCR].
Zhurnal mikrobiologii, epidemiologii i immunobiologii
2015 , Issue 6, Page(s) 35–42
Abstract: ... serogroup A, B, C and W DNA.: Materials and methods: Reference strains and 187 samples ... of clinical CSF samples: A--in 103 samples (55%), B--in 45 (24%), C--in 30 (16%), W--in 5 (3%). Only DNA ... A menincococci as well as DNA of 5 group W meningococci and 4 ungroupable was carried out.: Conclusion ...
Abstract | Aim: Development and testing of a real-time PCR method for detection of Neisseria meningitiis serogroup A, B, C and W DNA. Materials and 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. Conclusion: 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. |
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MeSH term(s) | Bacterial Capsules/genetics ; Bacterial Outer Membrane Proteins/genetics ; DNA, Bacterial/genetics ; Gene Expression ; Genetic Loci ; Genome, Bacterial ; Genotype ; Humans ; Meningitis, Meningococcal/cerebrospinal fluid ; Meningitis, Meningococcal/microbiology ; Multilocus Sequence Typing ; Multiplex Polymerase Chain Reaction ; Neisseria meningitidis/classification ; Neisseria meningitidis/genetics ; Neisseria meningitidis/isolation & purification ; Porins/genetics ; Serogroup ; Serotyping |
Chemical Substances | Bacterial Outer Membrane Proteins ; DNA, Bacterial ; FrpB protein, bacteria ; Porins ; porin protein, Neisseria |
Language | Russian |
Publishing date | 2015-04-01 |
Publishing country | Russia (Federation) |
Document type | English Abstract ; Journal Article |
ZDB-ID | 218354-7 |
ISSN | 0372-9311 ; 0049-8726 ; 0372-8714 |
ISSN | 0372-9311 ; 0049-8726 ; 0372-8714 |
Database | MEDical Literature Analysis and Retrieval System OnLINE |
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