title: Incorporation of real-time PCR into routine public health surveillance of culture negative bacterial meningitis in São Paulo, Brazil.
abstract: Real-time (RT)-PCR increases diagnostic yield for bacterial meningitis and is ideal for incorporation into routine surveillance in a developing country. We validated a multiplex RT-PCR assay for Streptococcus pneumoniae, Neisseria meningitidis, and Haemophilus influenzae in Brazil. Risk factors for being culture-negative, RT-PCR positive were determined. The sensitivity of RT-PCR in cerebrospinal fluid (CSF) was 100% (95% confidence limits, 96.0%-100%) for N. meningitidis, 97.8% (85.5%-99.9%) for S. pneumoniae, and 66.7% (9.4%-99.2%) for H. influenzae. Specificity ranged from 98.9% to 100%. Addition of RT-PCR to routine microbiologic methods increased the yield for detection of S. pneumoniae, N. meningitidis, and H. influenzae cases by 52%, 85%, and 20%, respectively. The main risk factor for being culture negative and RT-PCR positive was presence of antibiotic in CSF (odds ratio 12.2, 95% CI 5.9-25.0). RT-PCR using CSF was highly sensitive and specific and substantially added to measures of meningitis disease burden when incorporated into routine public health surveillance in Brazil.
authors: Division of Medical Biology, Department of Immunology, Instituto Adolfo Lutz, São Paulo, Brazil. ctsacchi@gmail.com
authors: Claudio T
authors: CT
authors: Sacchi
authors: Lucila O
authors: LO
authors: Fukasawa
authors: Maria G
authors: MG
authors: Gonçalves
authors: Maristela M
authors: MM
authors: Salgado
authors: Kathleen A
authors: KA
authors: Shutt
authors: Telma R
authors: TR
authors: Carvalhanas
authors: Ana F
authors: AF
authors: Ribeiro
authors: Brigina
authors: B
authors: Kemp
authors: Maria C O
authors: MC
authors: Gorla
authors: Ricardo K
authors: RK
authors: Albernaz
authors: Eneida G L
authors: EG
authors: Marques
authors: Angela
authors: A
authors: Cruciano
authors: Eliseu A
authors: EA
authors: Waldman
authors: M Cristina C
authors: MC
authors: Brandileone
authors: Lee H
authors: LH
authors: Harrison
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D015373:Bacterial Typing Techniques; D001938:Brazil; D002648:Child; D002675:Child, Preschool; D005260:Female; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D007958:Leukocyte Count; D008297:Male; D016920:Meningitis, Bacterial / Q000097:blood / Q000134:cerebrospinal fluid / Q000382:microbiology*; D008875:Middle Aged; D015999:Multivariate Analysis; D009345:Neisseria meningitidis / Q000145:classification / Q000235:genetics; D011159:Population Surveillance / Q000379:methods*; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012307:Risk Factors; D012680:Sensitivity and Specificity; D055815:Young Adult