Use of an automated DNA analysis system (DARAS (R)) for sequence-specific recognition of Neisseria meningitidis DNA

Objective To combine use of the polymerase chain reaction (PCR) for rapid d iagnosis of meningococcal meningitis with a novel automated detection syste m for sequence-specific recognition of PCR products. Methods DNA was extracted from cerebrospinal fluid (CSF) by a quick boil-ly sis method, followed by PCR with printers specific for Neisseria meningitid is. Sequence-specific recognition of N. meningitidis DNA was performed with an automated DNA analysis system (DARAS(R)) and the data were compared wit h results following agarose gel electrophoresis or conventional microbiolog ical culture. Results The DARAS(R) system had a sensitive detection limit of 10(2) mening ococci/mL with spiked samples, compared with a detection limit of 10(4) men ingococci/mL following agarose gel electrophoresis. When the system was use d to examine 74 CSF samples, the 19 CSF samples positive for N. meningitidi s by conventional microbiological methods were also all positive in the DAR AS(R) system and the 55 samples negative by DARAS(R) for meningococci were also negative by conventional microbiological methods. Conclusion The sensitivity and specificity of the DARAS(R) system makes it a useful tool for the diagnosis of meningococcal meningitis. The system is user-friendly, requires minimal hands-on time and generates data in an info rmative numerical format.