title: Diagnosis and species identification of mycobacterial infections by polymerase chain reaction-restriction fragment length polymorphism analysis of sterile body fluids.
abstract: OBJECTIVE
  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 PCR-RFLP 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.
authors: AIDS Research Institute, Yonsei University College of Medicine, Seodaemun-gu, Seoul, Korea.
authors: Cheong Ho
authors: CH
authors: Cho
authors: Sang Hoon
authors: SH
authors: Han
authors: Bum Sik
authors: BS
authors: Chin
authors: Suk Hoon
authors: SH
authors: Choi
authors: Han Sung
authors: HS
authors: Lee
authors: Chang Oh
authors: CO
authors: Kim
authors: Myung Soo
authors: MS
authors: Kim
authors: Jun Yong
authors: JY
authors: Choi
authors: Young Goo
authors: YG
authors: Song
authors: June Myung
authors: JM
authors: Kim
mesh_terms: D054458:Amplified Fragment Length Polymorphism Analysis; D001426:Bacterial Proteins / Q000235:genetics*; D015373:Bacterial Typing Techniques; D001826:Body Fluids / Q000382:microbiology*; D004269:DNA, Bacterial / Q000032:analysis*; D012321:DNA-Directed RNA Polymerases / Q000235:genetics*; D006801:Humans; D009161:Mycobacterium / Q000145:classification* / Q000235:genetics; D009164:Mycobacterium Infections / Q000175:diagnosis* / Q000382:microbiology; D012150:Polymorphism, Restriction Fragment Length
chemical_list: D001426:Bacterial Proteins; D004269:DNA, Bacterial; D012321:DNA-Directed RNA Polymerases