title: Assessment of a two-step nucleic acid amplification assay for detection of Neisseria meningitidis followed by capsular genogrouping.
abstract: Immediate prevention of meningococcal disease relies in part on the prompt treatment with antibiotics of household and other close contacts of cases; however intervention with effective vaccination relies on identification of serogroup-causing strains. Parenteral antibiotic for patient with suspected meningococcal disease before hospital admission is currently recommended. Laboratory standard methods are hindered by failure to detect bacteria by this medical approach to improve patient prognosis. We assessed two polymerase chain reaction (PCR) assays to detect (crgA) and define the serogroups (siaD, orf-2, and ctrA) of Neisseria meningitidis in 120 cerebrospinal fluid (CSF) samples from positive cases (culture or antigen detection or direct smear). The PCR sensitivity for the identification of N. meningitidis was 100% (95% confidence interval, CI, 96-100%) compared to a sensitivity of 46% for culture (95% CI 37-55%), 61% for latex agglutination test (95% CI 52-70%), and 68% for Gram stain (95% CI 59-76%); PCR specificity was 97% (95% CI 82-100%). PCR correctly identified the serogroups A, B, C, W135, Y, and X in CSF samples with a sensitivity of 88% (95% CI 80-93%); the primer sets were 100% specific. The introduction of PCR-based assays shall increase laboratory confirmed cases, consequently enhancing surveillance of meningococcal disease.
authors.affiliation: Departamento de Bacteriologia, Instituto Oswaldo Cruz-Fiocruz, Qv. Brasil 4365, 21040-900 Rio de Janeiro, RJ, Brazil.
authors.forename: Maria C
authors.initials: MC
authors.lastname: Rebelo
authors.forename: Renata F
authors.initials: RF
authors.lastname: Boente
authors.forename: Juliana de A
authors.initials: Jde A
authors.lastname: Matos
authors.forename: Cristina B
authors.initials: CB
authors.lastname: Hofer
authors.forename: David E
authors.initials: DE
authors.lastname: Barroso
mesh_terms: D000293:Adolescent; D000328:Adult; D001426:Bacterial Proteins / Q000235:genetics*; D002648:Child; D002675:Child, Preschool; D004269:DNA, Bacterial / Q000134:cerebrospinal fluid*; D005260:Female; D006801:Humans; D007223:Infant; D008297:Male; D008585:Meningitis, Meningococcal / Q000134:cerebrospinal fluid / Q000175:diagnosis* / Q000382:microbiology; D008875:Middle Aged; D009345:Neisseria meningitidis / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D016133:Polymerase Chain Reaction; D011446:Prospective Studies; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012703:Serotyping; D014157:Transcription Factors / Q000235:genetics*
chemical_list: D001426:Bacterial Proteins; C406769:CrgA protein, Neisseria meningitidis; D004269:DNA, Bacterial; D014157:Transcription Factors