Background Meningitis is the most dangerous disease in children and remained irreversible mental disorders. H. influenzae is a fastidious bacteria and may be under detected because of inadequate techniques for isolation or overuse of antibiotics before with recovery of causative agents in bacterial meningitis.in present study two methods, culture and molecular diagnosis (PCR) apply for isolating H.I from CSF.
Methods DNA was extracted from CSF and probed for the presence of Hib DNA with PCR assay with primer derived from the sequences encoding a capsulation-associated protein; a protein most probably involved in the intracellular transportation of the capsular polysaccharide, and would be expected to react only with capsulate H. influenzae strains. Primers sequencing were:
Primer 1: 5′- CGT TTG TAT GAT GTT GAT CCA GAC T
Primer 2: 5′- TGT CCA TGT CTT CAA AAT GAT G
Results Two hundred three cerebrospinal fluid (CSF) samples collected consecutively from children(less than 5 years) suffering from meningitis were investigated by PCR. There were all the cases of clinical meningitis admitted to three children hospitals in 18 months duration period.
Discussion Two hundred CSF samples were investigated by PCR. Seven samples were positive by PCR method (5 samples were culture positive and 2 samples were culture negative for Haemophilus influenza). Haemopjilus influenza type b is a agent 17.1% of bacterial meningitis in children surveyed.