All issues > Volume 40(9); 1997

Original Article

J Korean Pediatr Soc. 1997;40(9):1232-1241. Published online September 15, 1997.

Rapid Identification of Mycoplasma Pneumoniae, Mycobacterium Tuberculosis and Staphylococcus Aureus in Pleural Fluid by PCR

Chang Wan CW Kim¹, Byung Moon BM Ahn¹, Eun Ryoung ER Kim¹, Il Su IS Kim², Yung Seuk YS Pak³, Sang Chul SC Sung³

¹Department of Pediatrics, Sung-Ae General Hospital, Seoul, Korea
²Sung-Ae Genentic Engeenearing Research Institute
³Department of Molecular Biology, Seoul Medical Science Institute, Seoul, Korea

Abstract

Purpose
: Pleural effusions may develop during the course of bacterial pneumonia. The aim of this study was to evaluate the significance of the polymerase chain reaction(PCR)
method
for detection of M ycoplasma pneumoniae, M ycobaterium tuberculosis and Staphylococcus aureus from pleural fluid.
Methods
: Total 12 samples were obtained from pleural fluid; 2 samples from children with Mycoplasma pneumonia, 5 samples from adults with tuberculous pleurisy, and 5 samples from sterile pleural fluid seeded artificially with staphylococcus aureus. The primers used for our PCR were prepared to amplify M . pneumonia-specific MP5 gene, M. tuberculosis-specific IS6110 gene, and S. aurus-specific femA and mecA gene. The amplified PCR products were detected by ethidium bromide-stained agarose gel electrophoresis.
Results
: A total of 12 pleural fluid samples were tested by nested PCR using the specific primer set. We could amplify MP5 gene in 2 samples, IS6110 gene in 5 samples, mecA gene in 3 samples, and femA gene in 5 samples. These PCR data were correlated with serolological data, microbiological data and methicillin-sensitivity test result. There were no false-positive results due to cross-contaminating DNA between these 3 organisms.
Conclusions
: We conclude that enzymatic amplification of specific gene from pleural fluid might be useful to diagnose the infectious pleural effusion by M ycoplasma pneumoniae, M ycobacterum tuberculosis or Staphylococcus aureus.

Keywords :M ycoplasma pneumoniae, M ycobacterium tuberculosis, Staphylococcus aureus, Pleural effusion, Polymerase chain reaction