Diagnosis and species identification of mycobacterial infections by polymerase chain reaction-restriction fragment length polymorphism analysis of sterile body fluids.
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.
Cho
Cheong Ho
CH
AIDS Research Institute, Yonsei University College of Medicine, Seodaemun-gu, Seoul, Korea.
Han
Sang Hoon
SH
Chin
Bum Sik
BS
Choi
Suk Hoon
SH
Lee
Han Sung
HS
Kim
Chang Oh
CO
Kim
Myung Soo
MS
Choi
Jun Yong
JY
Song
Young Goo
YG
Kim
June Myung
JM
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
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D012321:DNA-Directed RNA Polymerases