PCR-ENZYME IMMUNOASSAY FOR DETECTION OF STREPTOCOCCUS-PNEUMONIAE DNA IN CEREBROSPINAL-FLUID SAMPLES FROM PATIENTS WITH CULTURE-NEGATIVE MENINGITIS

A PCR-based assay was developed to amplify a conserved region of the p neumococcal autolysin gene. The amplified product was labelled with di goxigenin-labelled dUTP and was detected with a biotin-labelled probe in an enzyme immunoassay (EW). The assay was initially tested with sus pensions of various serotypes of Streptococcus pneumoniae and other gr am-positive and gram-negative bacteria and was then applied to cere br ospinal fluid (CSF) specimens from patients with meningitis and those with other neurological disorders. The assay detected all the serotype s of S. pneumoniae tested, whereas all the other bacterial strains tes ted were negative. Seven of the 8 CSF specimens positive for pneumococ cus by culture or latex agglutination (LA) were positive by PCR-EIA wh ereas all 10 specimens positive for other organisms were negative. Amo ng 11 patients with clinically diagnosed meningitis but with negative culture and LA results, 5 were positive by PCR EIA. The assay was nega tive for all but one patient without meningitis; it was positive with the CSF from a child with immunodeficiency and pneumococcal abscesses on the scalp. PCR-EW is a useful tool for the diagnosis of meningitis, especially when culture and LA are negative because of prior antibiot ic treatment.