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

• Published in: European journal of clinical microbiology & infectious diseases Vol. 19; no. 1; pp. 47 - 50
• Main Authors: BARAN, J. JR, RIEDERER, K. M, KHATIB, R
• Format: Journal Article
• Language: English
• Published: Berlin Springer 01.02.2000; Springer Nature B.V
• Subjects: Bacterial diseases; Bacterial diseases of the nervous system. Bacterial myositis; Biological and medical sciences; Cerebrospinal fluid; Cerebrospinal Fluid - microbiology; Colony Count, Microbial; Culture Media; DNA, Bacterial - analysis; Evaluation Studies as Topic; Human bacterial diseases; Humans; Immunoassay; Infectious diseases; Laboratories; Medical sciences; Meningitis; Mycobacterium tuberculosis; Mycobacterium tuberculosis - genetics; Mycobacterium tuberculosis - growth & development; Mycobacterium tuberculosis - isolation & purification; Polymerase chain reaction; Polymerase Chain Reaction - methods; Radiometry; Sensitivity and Specificity; Tuberculosis; Tuberculosis, Meningeal - diagnosis; Tuberculosis, Meningeal - microbiology; United States > US; Detroit Michigan; Human; Nervous system diseases; Mycobacterial infection; Cerebrospinal fluid; Method; Infection; Polymerase chain reaction; Microbiological investigation; Tuberculosis; Mycobacterium tuberculosis; Mycobacteriales; Central nervous system disease; Limitating dilution; Detection threshold; Bacteriosis; Mycobacteriaceae; Bacteria; Meningitis; Actinomycetes; Diagnosis; Molecular biology
• ISSN: 0934-9723; 1435-4373
• DOI: 10.1007/s100960050009

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).