[Optimization of real-time multiplex polymerase chain reaction for the diagnosis of acute bacterial meningitis and Neisseria meningitidis serogrouping].

In this study, three Rt-multiplex PCR assays were optimized and standardized for the detection of N.meningitidis, H.influenzae and S.pneumoniae (NHS mix) and for the serogrouping of N.meningitidis (BCY mix and AWX mix) in our laboratory. Sensitivity of the Rt-multiplex PCR method was detected by reference strains and simulation studies. Forty three samples (41 cerebrospinal fluid (CSF), 1 serum and 1 clinical isolate) with the suspicion of acute bacterial meningitis and six nasopharyngeal isolates sent to National Reference Laboratory for Molecular Microbiology between July 2012-May 2014 were included in the present study. All samples were examined with the Rt-multiplex PCR methods. Clinical isolates were studied by both conventional and Rt-multiplex PCR methods. Oxidase, catalase test, Gram stain, API-NH and agglutination tests with specific antisera were performed in the National Respiratory Pathogens Reference Laboratory. The detection limit of the method, which was optimized and standardized in our laboratory was determined as 102 cfu/ml. The CT (threshold cycle) value in this dilution was detected approximately as 35. N.meningitidis was detected in 14 of the 41 CSF samples by the NHS mix. However, only 10 of the positive samples could be typed with BCY mix and AWX mix. Eight (80%) of them were serogroup B, one of each was (10%) serogroup A and serogroup W135, respectively. All the isolates (six nasopharyngeal and one clinical specimen) were identified as N.meningitidis s...
Source: Mikrobiyoloji Bulteni - Category: Microbiology Tags: Mikrobiyol Bul Source Type: research