A prospective study (from August 2006 to April 2007) was carried out with 214 cerebrospinal fluid samples with suspicion of bacterial meningitis. The aim of the study was to assess the effectiveness of the simultaneous detection of Neisseria meningitidis, Streptococcus sp. and Haemophilus influenzae using seminested polymerase chain reaction strategy. Among the 214 samples tested by both PCR and culture, the overall confirmation rate was 64% for PCR and 40.1% for culture (P = 2 x 10⁻⁶). Taking culture method as the standard reference, the overall sensitivity of PCR was 98.8% and specificity, 59.4%. The sensitivity of PCR was 100, 97.3 and 100% respectively for N. meningitidis, Streptococcus sp. and H. influenzae with respective specificities of 70, 93.2 and 97.2%. In conclusion, the seminested PCR strategy is a sensitive method and it can be implemented in the reference public health laboratories for an exhaustive microbiological surveillance of bacterial meningitis.