Journal List > Korean J Lab Med > v.26(3) > 1011307

Park, Heo, Park, Song, Lee, Lee, and Kim: Isolation of Nontuberculous Mycobacteria Using Polymerase Chain Reaction-Restriction Fragment Length Polymorphism

Abstract

Background

The isolation rate of nontuberculous mycobacteria (NTM) in clinical laboratories and the incidence of NTM infections are on the increase recently in Korea, but there have been only a few studies that reveal the general aspect of NTM isolation or species distribution. Therefore, this study was performed to examine the isolation rate of NTM, species identification, and the clinical significance of mycobacterial cultures.

Methods

From August 2004 to May 2005, we examined mycobacterial isolates by AccuProbe test to differentiate NTM from Mycobacterium tuberculosis complex. NTM was then identified by polymerase chain reaction-restriction fragment length analysis (PCR-RFLP).

Results

A total of 6,742 specimens from 2,784 patients were requested for mycobacterial culture. Mycobacteria were isolated from 776 specimens (11.5%). The isolation rates of NTMs among the total culture positive specimens and culture positive sputum specimens were 24.4% (189/776) and 25.3% (169/667), respectively. Fourteen species of NTM identified in 172 of the 175 specimens tested included M. avium (39.0%), M. intracellulare (22.7%), and M. abscessus (19.8%).

Conclusions

Using AccuProbe tests and PCR-RFLP method for mycobacterial cultures processed in a clinical laboratory, we were able to identify NTMs to the species level. The isolation rate of NTM in this study was similar to that reported in past studies in Korea. In addition, we found that some of the NTMs isolated in this study could cause pulmonary diseases.

References

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Table 1.
Summaries of mycobacterial culture by specimen types
Type N of specimens (%)
Total Culture-positive MTB NTM
Sputum 5,261 (78.0) 667 (86.0) 498 (74.7) 169 (25.3)
Bronchial washing 469 (7.0) 52 (6.7) 38 (73.1) 14 (26.9)
CSF 294 (4.4) 5 (0.6) 3 (60.0) 2 (40.0)
Pleural fluid 254 (3.8) 19 (2.4) 19 (100.0)
Tissue 95 (1.4) 13 (1.7) 11 (84.6) 2 (15.4)
Ascites 66 (1.0) 1 (0.1) 1 (100.0)
Wound 46 (0.7) 4 (0.5) 3 (75.0) 1 (25.0)
Abscess 38 (0.6) 3 (0.4) 3 (100.0)
CPS 33 (0.5) 4 (0.5) 4 (100.0)
Pericardial fluid 12 (0.2) 2 (0.3) 2 (100.0)
Others 174 (2.6) 6 (0.8) 5 (83.3) 1 (16.7)
Total 6,742 (100.0) 776 (100.0) 587 (75.6) 189 (24.4)

Abbreviations: MTB, M. tuberculosis complex; NTM, nontuberculous mycobacteria; CSF, cerebrospinal fluid; CPS, cerebroperitoneal shunt.

Table 2.
Characteristics of isolated nontuberculous mycobacteria species
Species name N of specimens (%)
N of patients (%)
Total Respiratory Total Acceptable* Pulmonary Ds.
M. avium complex 106 (62.2) 106 (63.9) 70 (64.8) 45 (78.9) 13 (68.4)
M. avium 68 (39.5) 68 (41.0) 49 (45.4) 29 (50.9) 6 (31.6)
M. intracellulare (type I, II) 39 (22.7) 39 (23.5) 21 (19.4) 16 (28.1) 7 (36.8)
M. abscessus 33 (19.2) 30 (18.1) 16 (14.8) 11 (19.3) 6 (31.6)
M. fortuitum 7 (4.1) 7 (4.2) 6 (5.6)
M. septicum 5 (2.9) 5 (3.0) 4 (3.7)
M. nonchromogenicum 4 (2.3) 4 (2.4) 3 (2.8)
M. asiaticum 3 (1.7) 3 (1.8) 2 (1.9)
M. mucogenicum 3 (1.7) 1 (0.6) 3 (2.8)
M. scrofulaceum 3 (1.7) 3 (1.8) 1 (0.9)
M. shimoidi 2 (1.2) 2 (1.2) 2 (1.9)
M. kansasii 2 (1.2) 2 (1.2) 2 (1.9) 1 (1.8)
M. gordonae 1 (0.6) 1 (0.6) 1 (0.9)
M. peregrinum 1 (0.6) 1 (0.6) 1 (0.9)
M. pulveris 1 (0.6) 1 (0.9)
Total 172 166 108 57 19
  (100.0) (100.0) (100.0) (100.0) (100.0)

* Patients that agree with specimen requirements defined by ATS guideline;

NTM pulmonary diseases diagnosed by ATS guideline.

Abbreviation: ATS, American Thoracic Society.

Table 3.
Comparison of isolation rates of nontuberculous mycobacteria species in Korea
Species names This study [N, %] [7] [N, %] [12] [N, %]
M. avium complex 106 (62.2) 138 (42.1) 34 (29.0)
M. avium 68 (39.5) 62 (18.9) 10 (8.5)
M. intracellulare (type I, II) 39 (22.7) 76 (23.2) 24 (20.5)
M. abscessus 33 (19.2) 37 (11.3) 4 (3.4)
M. fortuitum complex 7 (4.1) 64 (19.5)* 1 (0.9)
M. septicum 5 (2.9)
M. nonchromogenicum 4 (2.3)
M. asiaticum 3 (1.7)
M. mucogenicum 3 (1.7) 3 (0.9)
M. scrofulaceum 3 (1.7)
M. shimoidi 2 (1.2)
M. kansasii 2 (1.2) 13 (4.0) 33 (28.2)
M. gordonae 1 (0.6) 33 (10.1) 31 (26.5)
M. peregrinum 1 (0.6)   2 (1.7)
M. pulveris 1 (0.6)
M. terrae complex 28 (8.5)
M. chelonae 7 (2.1)
M. celatum 2 (0.6)
Others 3 (1.2) 12 (10.3)
Total 172 (100.0) 328 (100.0) 117 (100.0)

* Fifteen of forty-one M. fortuitum complex were proved to be M. peregrinum via further analysis;

each 1 case of M. gastri, M. szulgai, M. violacefusca;

1 M. marinum and 11 unclassified NTM.

Table 4.
Clinical characteristics of NTM pulmonary diseases by ATS guidelines
No. case Age (year) Sex Culture specimen Clinical symptoms Underlying diseases Radiologic findings of chest Isolated NTM
1 56 F bronchial washing cough, sputum multifocal bronchiectasis M. avium
2 76 F sputum cough old Tb centrilobular nodules M. avium
3 47 F sputum chest pain old Tb centrilobular nodules M. avium
4 29 F sputum hemoptysis, lymph node enlargement multifocal bronchiectasis M. avium
5 66 M sputum dyspnea, hemoptysis old Tb multifocal bronchiectasis M. avium
6 65 F sputum fever, cough diabetes, hypertension multifocal bronchiectasis M. avium
7 62 F sputum fever multiple cavities M. intracellulare
8 64 M sputum hemoptysis asthma multifocal consolidation & bronchiectasis M. intracellulare
9 78 F sputum cough, sputum old Tb bronchovascular infiltration M. intracellulare
10 57 M sputum cough, sputum multiple nodules M. intracellulare
11 70 M sputum fever old Tb multifocal patchy opacity M. intracellulare
12 76 M sputum hemoptysis multifocal bronchiectasis M. intracellulare
13 50 F sputum hemoptysis multifocal bronchiectasis M. intracellulare
14 78 F sputum cough, sputum, fever centrilobular nodules M. abscessus
15 45 M sputum cough MDS multiple nodules M. abscessus
16 47 F bronchial washing cough optic neuritis centrilobular nodule & bronchiectasis M. abscessus
17 76 M sputum fatigue old Tb multiple nodular consolidation M. abscessus
18 64 F sputum dyspnea asthma multifocal bronchiectasis M. abscessus
19 70 M sputum cough, sputum, fatigue optic neuritis nodular patchy opacity M. abscessus

Abbreviations: NTM, nontuberculous mycobacteria; ATS, American Thoracic Society; Tb, tuberculosis; MDS, myelodysplastic syndrome.

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