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Neisseria meningitidis PorA variable regions: rapid detection of P1·7 and P1·19 variants by PCR

2007; Oxford University Press; Volume: 45; Issue: 4 Linguagem: Inglês

10.1111/j.1472-765x.2007.02205.x

1472-765X

Ivano de Filippis, Claudia Ferreira de Andrade, Antônio Eugênio Castro Cardoso de Almeida, Maysa Mandetta Clementino, Cristiano Augusto Coelho Fernandes, Manuela Carvalho, Ana Carolina Paulo Vicente,

Pneumonia and Respiratory Infections

Aim: 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. Methods and 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. Significance and Impact of the Study: 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.