기관지 검체에서의 마이코박테리아 검출 및 동정을 위한 BacT/ALERT 3D System과 PCR-RFLP 병용 = Evaluation of Combined Use of BacT/ALERT 3D Liquid Culture System and PCR-RFLP for Detection and Identification of Mycobacteria from Bronchial Specimens

다국어 초록 (Multilingual Abstract)

Background: We evaluated BacT/ALERT 3D liquid
culture system (bioMérieux, USA) and PCR-restriction
fragment length polymorphism (RFLP) for recovery
and direct identification of mycobacteria, and the
results were compared with a conventional culture
system using an egg-based solid medium.
Methods: A total of 3,037 bronchial specimens (2,309
bronchial washing fluids and 728 bronchoalveolar
lavages) were collected. Decontaminated specimens
were inoculated to both BacT/ALERT MP liquid media
and Ogawa solid media (3%, Shinyang, Korea).
Recovery rate and detection time were compared between
the two systems. Liquid media from positive
cultures were centrifuged and the pellets were tested
for direct identification of mycobacteria by PCR-RFLP
using Myco-ID (M&D Inc., Korea).
Results: A total of 518 isolates, including 215 M. tuberculosis
(MTB) and 303 non-tuberculosis mycobacteria
(NTM), were recovered. The liquid media detected
492 isolates (16.2%), including 195 MTB and
297 NTM), whereas the solid media detected 416
isolates (13.7%), including 187 MTB and 229 NTM
(P＜0.001); 102 isolates (28 MTB and 74 NTM) were
recovered only by the liquid media, while 26 (20
MTB and 6 NTM) isolates were recovered only by
the solid media. The mean time to detection was
18.1 days by the liquid media and 29.3 days by the
solid media (P＜0.001). The overall time to species
identification from inoculation was 21.8 days. Direct
PCR-RFLP from the liquid media identified 39.1% of
MTB, 6.3% of M. avium, 19.05 of M. abscessus, and
12.6% of M. intracellulare respectively.
Conclusion: Combined use of a liquid culture system
and PCR-RFLP improved the recovery rate and shortened
the detection time. However, solid media is
still necessary to maximize the diagnostic efficiency.

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