Korean J Intern Med.
2009 Jun;24(2):135-138. 10.3904/kjim.2009.24.2.135.
Diagnosis and Species Identification of Mycobacterial Infections by Polymerase Chain Reaction-Restriction Fragment Length Polymorphism Analysis of Sterile Body Fluids
- Affiliations
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- 1AIDS Research Institute, Yonsei University College of Medicine, Seoul, Korea.
- 2Department of Internal Medicine, Yonsei University College of Medicine, Seoul, Korea. mkim@yuhs.ac
- 3Brain Korea 21 Project for Medical Sciences, Yonsei University College of Medicine, Seoul, Korea.
- KMID: 1717964
- DOI: http://doi.org/10.3904/kjim.2009.24.2.135
Abstract
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BACKGROUND/AIMS: The development of effective, accurate, and rapid diagnostic methods for Mycobacterium infection and mycobacterial species identification is required. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) is an easy, rapid and inexpensive technique for identifying Mycobacterium spp.
METHODS
We performed PCR-RFLP to detect and identify Mycobacterium spp. from 10 sterile body fluids, including ascites, cerebrospinal fluid, pleural fluid, synovial fluid, and peritoneal dialysis fluid. Clinical samples were collected from patients with diagnoses of definite, probable or suspected mycobacterial infection. The conserved RNA polymerase genes of Mycobacterium spp. were amplified by PCR.
RESULTS
The amplified 360-bp region of rpoB was digested with the restriction enzyme MspI or HaeIII. The PCRRFLP results for the clinical samples were identical to those for M. tuberculosis, M. fortuitum, M. intracellulare, and M. avium. In addition, the results of the PCR-RFLP were identical to those obtained by DNA sequencing.
CONCLUSIONS
PCR-RFLP analysis of sterile body fluids may be a useful method for the diagnosis of mycobacterial infections and for the differentiation of mycobacterial species.
Keyword
MeSH Terms
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*Amplified Fragment Length Polymorphism Analysis
Bacterial Proteins/*genetics
*Bacterial Typing Techniques
Body Fluids/*microbiology
DNA, Bacterial/*analysis
DNA-Directed RNA Polymerases/*genetics
Humans
Mycobacterium/*classification/genetics
Mycobacterium Infections/*diagnosis/microbiology
*Polymorphism, Restriction Fragment Length
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