Direct PCR assay for detection of Neisseria meningitidis in human cerebrospinal fluid.
A PCR amplification was performed to detect Neisseria meningitidis insertion sequence 1106 (IS-1106) in the human CerebroSpinal Fluid (CSF) in cases of meningitis. The study included 27 CSF samples from suspected meningitis patients. Although the inflammatory response in most of the samples was slightly increased, the results showed that 7 (26%) and 8 (30%) CSF samples were diagnosed as meningococcal meningitis by Gram staining and by culture, respectively. The primers of the IS-1106 were used for direct diagnosis of N. meningitidis in the human spinal fluid after a minor treatment of the CSF samples. The sample was diagnosed as meningococcal meningitis, if a DNA band of about 600 bp was detected in the ethidium bromide-stained agarose gel. The 27 CSF samples were analyzed in a random manner. Of these, 18 samples including the Gram staining- and culture-positive samples were also positive in PCR amplification. However, a CSF sample, which was diagnosed to be meningococcal meningitis in culture was negative in both Gram staining and PCR analysis. The specificity of the IS-1106 primers was determined to be 95%, with 100% sensitivity in comparison to Gram staining and culture. The primers were sensitive to 10 pg or more of meningococcal DNA. In addition, the PCR amplification showed high predictive values (89 and 100%) in diagnosing meningitis in patients that were negative and positive responders when tested by culture and by Gram staining. In conclusion, the PCR amplification of IS-1106 of N. meningitidis is specific and sensitive to both culture-positive and -negative meningococcal meningitis. Hence, PCR assay is highly recommended for use in a rapid diagnosis of suspected meningitis patients.
Abdel-Salam
H A
HA
Department of Microbiology, Faculty of Pharmacy, Zagazig University, Egypt.
D001483:Base Sequence; D002555:Cerebrospinal Fluid / Q000382:microbiology*; D017931:DNA Primers / Q000235:genetics; D006801:Humans; D008585:Meningitis, Meningococcal / Q000134:cerebrospinal fluid / Q000175:diagnosis / Q000382:microbiology; D009345:Neisseria meningitidis / Q000235:genetics* / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods*
D017931:DNA Primers