The diagnostic use of the polymerase chain reaction for the detection of Mycobacterium tuberculosis

• Published in: Pathology Vol. 26; no. 4; p. 482
• Main Authors: Weekes, K M, Pearse, M J, Sievers, A, Ross, B C, d'Apice, A J
• Format: Journal Article
• Language: English
• Published: England 01-10-1994
• Subjects: Base Sequence; DNA, Bacterial - isolation & purification; Evaluation Studies as Topic; Humans; Molecular Sequence Data; Mycobacterium tuberculosis - genetics; Mycobacterium tuberculosis - isolation & purification; Nucleic Acid Synthesis Inhibitors; Polymerase Chain Reaction; Sensitivity and Specificity; Sputum - microbiology; Taq Polymerase
• ISSN: 0031-3025
• DOI: 10.1080/00313029400169232

Detection of Mycobacterium tuberculosis by microscopy is difficult in specimens containing fewer than 10(4) bacteria/mL and growth in culture can take up to 6 wks. In this study the Polymerase Chain Reaction (PCR) was investigated as a rapid diagnostic technique for the detection of M. tuberculosis. The presence of DNA polymerase inhibitors in sputum specimens poses a potentially serious problem as false negative results can occur. In this study polymerase inhibitors were detected by inclusion of an internal plasmid control in each test. DNA from specimens in which the internal control failed to amplify was purified with a DNA binding matrix before retesting by PCR. A total of 169 sputum specimens were examined and 4 were found to have inhibitors. The correlation between detection of M. tuberculosis by PCR with a combination of culture, Ziehl-Neelsen (ZN) staining and patient history was 97.6%. This study confirms that PCR offers a more sensitive and rapid alternative for the detection of M. tuberculosis to ZN staining and culture, with results being available within 24 hrs of a specimen being received in the laboratory.