Limits of detection of Mycobacterium tuberculosis in spiked cerebrospinal fluid using the polymerase chain reaction in tuberculous meningitis.

Abstract

The limit of detection of Mycobacterium tuberculosis in spiked cerebrospinal fluid (CSF) using polymerase chain reaction (PCR) was compared to that of a radiometric liquid culture. Serial dilutions of clinical isolates of Mycobacterium tuberculosis were prepared in CSF (n=3) or broth (n=11) with estimated concentrations of 0-550 cfu/ml. Each dilution was examined concurrently by PCR and radiometric culture. PCR and radiometric culture detected Mycobacterium tuberculosis DNA in all dilutions with an estimated 2 cfu/ml in the CSF. At lower concentrations (estimated <2 cfu/ml), PCR and radiometric culture were positive in three of five (60%) and five of five (100%) CSF samples, respectively. In comparison to PCR in broth dilutions, no evidence of inhibition or interference was noted. These results imply that PCR can provide a rapid and reliable diagnosis of tuberculous meningitis, although there is a potential for false-negative results to occur in samples containing very few organisms ( < 2 cfu/ml).