Tuberculosis and Respiratory Diseases

The Korean Academy of Tuberculosis and Respiratory Diseases (대한결핵및호흡기학회)
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Detection of Mycobacterium tuberculosis DNA by PCR in Peripheral Blood of Patients with Pulmonary Tuberculosis

폐결핵 환자의 말초 혈액에서 중합효소연쇄반응을 이용한 결핵균 DNA의 검출

  • Hong, Yoon Ki (Division of Pulmonary and Critical Care Medicine, University of Ulsan College of Medicine, Asan Medical Center) ;
  • Jo, Kyung Uk (Division of Pulmonary and Critical Care Medicine, University of Ulsan College of Medicine, Asan Medical Center) ;
  • Lee, Hyeyoung (Department of Biomedical Laboratory Science, College of Health Sciences, Yonsei University) ;
  • Kim, Mi-Na (Department of Laboratory Medicine, University of Ulsan College of Medicine, Asan Medical Center) ;
  • Sung, Heungsup (Department of Laboratory Medicine, University of Ulsan College of Medicine, Asan Medical Center) ;
  • Oh, Yeon-Mok (Division of Pulmonary and Critical Care Medicine, University of Ulsan College of Medicine, Asan Medical Center) ;
  • Lee, Sang Do (Division of Pulmonary and Critical Care Medicine, University of Ulsan College of Medicine, Asan Medical Center) ;
  • Kim, Woo Sung (Division of Pulmonary and Critical Care Medicine, University of Ulsan College of Medicine, Asan Medical Center) ;
  • Kim, Dong Soon (Division of Pulmonary and Critical Care Medicine, University of Ulsan College of Medicine, Asan Medical Center) ;
  • Kim, Won Dong (Division of Pulmonary and Critical Care Medicine, University of Ulsan College of Medicine, Asan Medical Center) ;
  • Shim, Tae Sun (Division of Pulmonary and Critical Care Medicine, University of Ulsan College of Medicine, Asan Medical Center)
  • 홍윤기 (울산의대 서울아산병원 호흡기내과) ;
  • 조경욱 (울산의대 서울아산병원 호흡기내과) ;
  • 이혜영 (연세대학교 원주의과대학 보건과학대학 임상병리학과) ;
  • 김미나 (울산의대 서울아산병원 진단검사의학과) ;
  • 성흥섭 (울산의대 서울아산병원 진단검사의학과) ;
  • 오연목 (울산의대 서울아산병원 호흡기내과) ;
  • 이상도 (울산의대 서울아산병원 호흡기내과) ;
  • 김우성 (울산의대 서울아산병원 호흡기내과) ;
  • 김동순 (울산의대 서울아산병원 호흡기내과) ;
  • 김원동 (울산의대 서울아산병원 호흡기내과) ;
  • 심태선 (울산의대 서울아산병원 호흡기내과)
  • Received : 2007.09.13
  • Accepted : 2007.10.09
  • Published : 2007.10.30
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Abstract

Background: Although pulmonary tuberculosis (TB) is a respiratory disease, the presence of Mycobacterium tuberculosis (Mtb) DNA or Mtb itself has been reported in the peripheral blood (PB) of several patients with pulmonary TB. Additionally, it was recently announced that active pulmonary TB patients donated PB, and that this blood was then transfused to other individuals in Korea. Methods: Sixty-nine patients with bacteriologically-confirmed pulmonary TB (35), non-tuberculous mycobacterial (NTM) lung disease (6), and other lung diseases (28) were enrolled in this study, which was conducted to determine if Mtb DNA could be detected in the PB by PCR. In addition, 10 pulmonary TB patients with high-burden bacilli were also enrolled in this study for the culture of Mtb in PB. Results: PCR detected the presence of Mtb in 22.8% (8/35) of the pulmonary TB patients, in 16.7% (1/6) of the patients with NTM lung disease, and in none of the patients with other diseases (0%). In addition, no Mtb was cultured from the PB of the 10 pulmonary TB patients. Conclusion: Although Mtb DNA was detected in the PB of some patients with pulmonary TB, viable Mtb was not isolated from the PB of those patients, which indicates that patients that viable Mth may not be transmitted via trasfusion of blood of pulmonary TB patients.

연구배경: 결핵은 호흡기를 통한 전염성 질환이지만 말초혈액에서 결핵균 PCR이 양성이거나 결핵균이 배양된 보고가 일부 있었다. 이는 결핵의 진단에 있어서의 유용성과 헌혈을 통한 결핵의 전염 가능성의 두 가지 문제를 제기하게 되었고, 저자들은 상기 문제들에 대한 향후 연구방향의 기초자료로 사용하고자 연구를 시행하여 보았다. 방법: 속립성 결핵이 아닌 폐결핵, 비결핵항산균 폐질환, 폐암 및 폐렴 환자 69명을 대상으로 말초혈액에서 결핵균 PCR을 시행하였고, 결핵균이 많을 것으로 추정되는 10명의 폐결핵 환자의 말초혈액을 대상으로 결핵균 배양을 시행하였다. 결핵균 PCR은 nested PCR을 이용한 TB-taq (M&D, Korea)을 이용하였고 혈액배양에는 BACTEC Myco/F Lytic 혈액배양병(Becton Dickinson, Sparks, Md)을 사용하였다. 결과: 결핵균 PCR을 시행한 환자는 69명으로 각각 폐결핵 35명, 비결핵항산균 폐질환 6명, 폐암 20명, 폐렴 8명이었다. 폐렴이나 폐암 환자 28명 모두에서 PCR은 음성이었고, 비결핵항산균 폐질환 6명 중 1명(16.7%), 폐결핵 35명 중 8명(22.8%)에서 양성이었다. 혈액 배양은 폐결핵 10명 모두에서 음성이었다. 결론: 미만성 결핵이 동반되지 않은 폐결핵 환자의 말초혈액에서 결핵균의 DNA가 검출됨을 확인할 수 있었으나 균배양이 되지 않는 것으로 보아 살아있는 균이 존재하기 보다는 균의 일부 성분만 존재할 가능성을 제시하였다.

Keywords

References

  1. Fujiwara H, Kleinhenz ME, Wallis RS, Eliner JJ, Increased interleukin-l production and monocyte suppressor cell activity associated with human tuberculosis. Am Rev Respir Dis 1986;133:73-7 https://doi.org/10.1164/arrd.1986.133.1.73
  2. Schluger NW, Condos R, Lewis S, Rom WN. Amplification of DNA of Mycobacterium tuberculosis from peripheral blood of patients with pulmonary tuberculosis. Lancet 1994;344:232-3 https://doi.org/10.1016/S0140-6736(94)92999-8
  3. Folgueira L, Delgado R, Palenque E, Aguado JM, Noriega AR. Rapid diagnosis of Mycobacterium tuberculosis bacteremia by PCR. J Clin Mcrohiol 1996; 34:612-6
  4. Condos R, McClune A, Rom WN, Schluger NW, Peripheral blood based PCR assay to identify patients with active pulmonary tuberculosis. Lancet 1996; 347:1082-5 https://doi.org/10.1016/S0140-6736(96)90281-0
  5. Kolk AH, Kox LF, Kuijper S, Richter C. Detection of Mycobacterium tuberculosis in peripheral blood. Lancet 1944;344:694
  6. Schiller NB, Stein SC. Whole blood transfusion as a determinant of tuberculin sensitivity. Am Rev Respir Dis 1969;100:740-1
  7. Sansila A, Hongmanee P, Chuchottawom C, Rienthong S, Rienthong D, Palittapongarnpim P. Differentiation between Mycobacterium tuberculosis and Mycobacterium aviun by amplification of the 16S-23S ribosomal DNA spacer. J Clin Mcrohiol 1998;36:2399-403
  8. Ahmed N, Mohanty AK, Mukhopadhyay D, Batish VK, Grover S. PCR-based rapid detection of Mycobacterium tuberculosis in blood from immunocom-petent patients with pulmonary tuberculosis. J Clin Microbiol 1998;36:3004-6
  9. Kim GW, Lee JM, Kang MJ, Son JW, Lee SJ, Kim LC, et al. Clinical significance of PCR-based rapid detection of Mycobacterium tuberculosis DNA in peripheral blood. Tuberc Respir Dis 2001;50:699-603
  10. Tjhie JH, van Belle AF, Thssens-Kroon M, van Soollngen D. Misidentification and diagnostic delay caused by a false-positive amplified Mycobacterium tuberculosis Direct Test in an immunocompetent patient with a Mycobacterium celatum infection. J Clin Microbiol 2001;39:2311-2 https://doi.org/10.1128/JCM.39.6.2311-2312.2001
  11. Chedore P, Broukhanski G, Shainhouse Z, Jamieson F. False-positive amplified Mycobacterium tuberculosis Direct Test results for samples containing Mycobacterium leprae. J Clin Mcrohiol 2006;44:612-3 https://doi.org/10.1128/JCM.44.2.612-613.2006