title: Polymerase chain reaction, with sequencing, as a diagnostic tool in culture---negative bacterial meningitis.
abstract: OBJECTIVE: To evaluate the feasibility of using 16S rDNA universal primer PCR (followed by sequencing) and 65-kDa heat shock Mycobacterium tuberculosis protein gene PCR as a method to determine a bacterial etiology in culture---negative cerebrospinal fluid (CSF) samples. METHODS: One hundred and forty-nine CSF samples from 128 patients were processed. DNA was extracted from the CSF samples and amplified with the eubacterial 16S rDNA primers P11E and P13B, and with the 65-kDa heat shock protein gene mycobacterial primers. The amplicons were identified by sequencing and specific oligoprobe hybridization. RESULTS: Overall, a microbiological diagnosis was made in 11 of 125 ultimately culture-negative cases. The use of 65-kDa heat shock protein gene PCR was needed to improve the diagnosis of tuberculous meningitis; in four patients, prospectively studied, the outcome of antituberculous therapy could also be followed. CONCLUSIONS: In culture-negative bacterial meningitis it is possible to improve the microbiological diagnosis by use of 16S rDNA amplification and sequencing, together with amplification of a more specific gene in mycobacteria.
authors.affiliation: Libera Università, Campus Bio-Medico.
authors.forename: Giulia
authors.initials: G
authors.lastname: Lorino
authors.forename: Daniela
authors.initials: D
authors.lastname: Lilli
authors.forename: Daniela
authors.initials: D
authors.lastname: Rivanera
authors.forename: Paola
authors.initials: P
authors.lastname: Guarino
authors.forename: Silvia
authors.initials: S
authors.lastname: Angeletti
authors.forename: Giovanni
authors.initials: G
authors.lastname: Gherardi
authors.forename: Francesco
authors.initials: F
authors.lastname: Filadoro