Journal List > Korean J Clin Microbiol > v.12(1) > 1038182

Chung, Ki, Lee, and Lee: Evaluation of Combined Use of BacT/ALERT 3D Liquid Culture System and PCR-RFLP for Detection and Identification of Mycobacteria from Bronchial Specimens

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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Table 1.
Isolations of mycobacteria from 3,037 specimens
Method BacT/ALERT liquid media
Egg-based solid media Result MTB NTM Total
+ − 167 28 20 2,822 223 74 6 2,734 390 102 26 2,519
P-value 0.312 <0.001 <0.001

Abbreviations: MTB, M. tuberculosis; NTM, nontuberculosis mycobacteria.

Table 2.
Time (days) to detect mycobacteria
Method BacT/ALERT liquid media Egg-based solid media P-value
MTB
NTM
24.6
12.8
30.8
28.0
<0.001
<0.001
Total 18.1 29.3 <0.001

Abbreviations: See Table 1.

Table 3.
Isolation frequency of mycobacteria from bronchial specimen
No. of isolates (%)
M. tuberculosis
M. avium
M. abscessus
M. intracellulare
M. fortuitum
M. kansasii
M. szulgai
M. chelonae
M. gordonae
171 (39.1)
115 (26.3)
83 (19.0)
55 (12.6)
2 (0.5)
2 (0.5)
2 (0.5)
1 (0.2)
6 (1.4)
Total 437
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