Rapid and efficient detection of extra-pulmonary Mycobacterium tuberculosis by PCR analysis

Int J Tuberc Lung Dis. 2000 Apr;4(4):361-70.

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 extrapulmonary 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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adolescent
  • Adult
  • Aged
  • Ascitic Fluid / microbiology
  • Child
  • Child, Preschool
  • DNA, Bacterial / genetics*
  • Exudates and Transudates / microbiology
  • Female
  • Humans
  • Infant
  • Male
  • Mexico
  • Middle Aged
  • Mycobacterium tuberculosis / genetics*
  • Pleural Effusion / microbiology
  • Polymerase Chain Reaction / methods*
  • Prospective Studies
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Tuberculosis / blood
  • Tuberculosis / cerebrospinal fluid
  • Tuberculosis / diagnosis*
  • Tuberculosis / microbiology*
  • Tuberculosis / urine

Substances

  • DNA, Bacterial