Rapid and efficient detection of extra-pulmonary Mycobacterium tuberculosis by PCR analysis
Abstract:SETTING: The diagnosis of extra-pulmonary tuberculosis (EPTB) remains an important clinical problem, primarily because of the inadequate sensitivity of conventional bacteriologic methods for detecting Mycobacterium tuberculosis in extra-pulmonary specimens.
OBJECTIVE: To evaluate whether a IS6110-based polymerase chain reaction (PCR) method can be utilized to detect M. tuberculosis in non-pulmonary specimens.
DESIGN: Specimens from 286 Mexican patients with a presumptive clinical diagnosis of EPTB were prospectively examined by Ziehl-Neelsen staining, mycobacterial culture on Löwenstein-Jensen slants, and by PCR. The DNA for PCR was extracted by the buffer lysis method and phenol-guanidine thiocyanate-chloroform. Primers that amplify a 200 bp fragment from the insertion-like M. tuberculosis sequence element IS6110 were utilized.
RESULTS: Our results demonstrate that this PCR method is highly specific (100%) for identifying M. tuberculosis from a variety of specimens including cerebrospinal fluid (CSF), pleural fluid, ascitic fluid, pericardial fluid, urine, and lymph node exudate. Moreover, the sensitivity of PCR for detecting M. tuberculosis in CSF (94%), pleural fluid (94%), ascitic fluid and other extra-pulmonary specimens (93%) greatly exceeds the sensitivity of conventional smear and culture methods.
CONCLUSION: These results demonstrate that PCR can be a highly specific and sensitive aid in the detection of M. tuberculosis from extra-pulmonary specimens.
Document Type: Regular Paper
Affiliations: 1: Molecular Biology in Medicine, Hospital Civil de Belén, CUCS, University of Guadalajara, Mexico 2: Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, Maryland, USA
Publication date: 2000-04-01
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