Neisseria meningitidis PorA variable regions: rapid detection of P1.7 and P1.19 variants by PCR.
OBJECTIVE
Rapid characterization of variable region (VR)1 variants of the porA gene among invasive strains is crucial for outbreak management and epidemiology studies. Recent sequence analysis studies in Brazil showed that the VR1 P1.7 and P1.19 variants are highly prevalent, accounting for 68%, of the total number of VR1 variants characterized. The aim of this work is to develop a rapid polymerase chain reaction (PCR)-based method for genosubtyping Neisseria meningitidis by detection of porA variable regions P1.7 and P1.19.


RESULTS
PCR primers for the detection of porA VR1 P1.7 and P1.19 were designed and tested using 198 clinical N. meningitidis isolates that had been previously evaluated by porA sequencing. All 50 strains with VR1 P1.7 and all 65 strains with VR1 P1.19 were positively identified by the respective VR-specific PCR and no false-positive reactions occurred.


CONCLUSIONS
VR-specific PCR amplification accurately identified VR P1.7 and P1.19 strains.


CONCLUSIONS
To overcome the disadvantages of serosubtyping and sequencing for typing the porA VR1 segment of N. meningitidis, we developed a PCR-based method to rapidly and accurately detect VR1 P1.7 and P1.19 variants. This approach is highly specific and sensitive; moreover it may allow for genotype determination of culture-negative samples.
de Filippis
I
I
Microbiology Dept., National Institute for Quality Control of Health, Oswaldo Cruz Foundation, Rio de Janeiro, Brazil. ivano.defilippis@incqs.fiocruz.br
de Andrade
C F
CF
de Almeida
A E C C
AE
Clementino
M M
MM
Fernandes
C A C M
CA
de Carvalho
M L
ML
Vicente
A C P
AC
D015373:Bacterial Typing Techniques; D001938:Brazil; D017931:DNA Primers; D006801:Humans; D008589:Meningococcal Infections / Q000175:diagnosis / Q000382:microbiology*; D009345:Neisseria meningitidis / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction; D018272:Porins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*
D017931:DNA Primers; D018272:Porins; C057699:porin protein, Neisseria