실시간-중합효소연쇄반응을 이용한 결핵균의 검출
Detection of Mycobacterium tuberculosis complex Using Real-time Polymerase Chain Reaction
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Abstract
Background : For the detection of Mycobacterium tuberculosis complex (MTB), PCR is known to be sensitive, specific, and rapid compared to the conventional methods of acid-fast-bacilli (AFB) smear and culture. We evaluated a new approach for MTB detection using real-time PCR. Methods : The specificity of real-time PCR was evaluated using 20 MTB isolates and 37 nontuberculous mycobacteria (NTM) isolates identified by AccuProbe Mycobacterium tuberculosis complex colony identification test (Gen-Probe Inc., USA) and Myco-ID (M&D, Korea). One hundred sputum specimens (50 AFB smear-positive and 50 negative specimens) were analyzed using real-time PCR and Amplicor Mycobacterium tuberculosis test (Roche, Germany). The results of real-time PCR positives (55 samples) and negatives (598 samples) were analyzed by AFB smear and culture. Results : The real-time PCR assay accurately discriminated between MTB and NTM species. Realtime PCR and Amplicor test yielded the same results in 96.0% (96/100) of the sputum specimens tested. The sensitivity and specificity of real-time PCR based on AFB culture were 97.4% and 88.5%, respectively. Of the 55 real-time PCR positive specimens, 83.6% (46/55) were culture-positive, 30.9 % (17/55) were smear-positive, 52.7% (29/55) were smear-negative and culture-positive, and 14.5% (8/55) were both smear and culture-negative. Among the 598 real-time PCR negative specimens, 60 were not tested for AFB smear or culture and 10 were contaminated. Of the remaining 528 specimens, 478 (90.5%) were both smear and culture-negative and 39 (7.4%) were culture-positive. Conclusions : For the detection of MTB, real-time PCR was sensitive and specific and comparable to conventional methods. It can be used for rapid identification of M. tuberculosis in clinical laboratories. (Korean J Lab Med 2008;28:103-8)
참고문헌 (19)
- Global tuberculosis control: surveillance, planning, financing (WHO Report 2004), "World Health Organization", World Health Organization, ~ (2004)
- Dye C, "Raviglione MC. Consensus statement. Global burden of tuberculosis: estimated incidence, prevalence, and mortality by country", WHO Global Surveillance and Monitoring Project, 282, pp.677~686 (1999), JAMA
- 고원중, "항산균 도말양성 객담에서 비결핵성 마이코박테리아의 분리 비율", Tuberculosis and Respiratory Diseases, 54(1), pp.22~32 (2003), 대한결핵및호흡기학회
- 임재준, "비결핵성 마이코박테리아 폐질환의 진단과 치료", 대한의사협회지, 48(6)통권 : 545, pp.563~570 (2005), 대한의사협회
- Marin M, "Rapid direct detection of multiple rifampin and isoniazid resistance mutations inMycobacterium tuberculosis in respiratory samples by real-time PCR", Antimicrob Agents Chemother, 48, pp.4293~4300 (2004)
- Wright PW, "Sensitivity of fluorochrome microscopy for detection of Mycobacterium tuberculosis versus nontuberculous mycobacteria", J Clin Microbiol, 36, pp.1046~1049 (1998)
- 고원중, "폐결핵의 진단과 치료", Tuberculosis and Respiratory Diseases, 58(5), pp.438~451 (2005), 대한결핵및호흡기학회
- Lee JY, "Clinical significance of low-colony count scotochromogen nontuberculous mycobacteria", Tuberc Respir Dis, 59, pp.39~46 (2005)
- 이재승, "객담 항산균 도말 양성 환자에서 비결핵항산균 과의 감별을 위한 결핵균 중합효소연쇄반응 검사의 유용성", Tuberculosis and Respiratory Diseases, 58(5), pp.452~458 (2005), 대한결핵및호흡기학회
- Choi YM, "Detection of Mycobacterium tuberculosis in sputum by using polymerase chain reaction", Korean J Clin Microbiol, 2, pp.144~151 (1999)
- Kim SR, "Clinical performance of the amplified Mycobacterium tuberculosis direct testrr for the detection of mycobacterium tuberculosis in non-respiratory specimens", Korean J Clin Pathol, 19, pp.315~319 (1999)
- Lee H, "Species identification of mycobacteria by PCR-restriction fragment length polymorphism of the rpoB gene", J Clin Microbiol, 38, pp.2966~2971 (2000)
- "Diagnostic standards and classification of tuberculosis in adults and children. This official statement of the American thoracic society and the centers for disease control and prevention was adopted by the ATS board of directors, July 1999. This statement was endorsed by the council of the infectious disease society of America, September 1999. Am J Respir Crit Care Med 2000;161:1376-95", ~ (1999)
- 박철민, "중합효소연쇄반응-제한절편길이다형성을 이용한 비결핵항산균의 분리", 대한진단검사의학회지, 26(3)통권 : 97, pp.161~167 (2006), 대한진단검사의학회
- Broccolo F, "Rapid diagnosis of mycobacterial infections and quantitation of Mycobacterium tuberculosis load by two real-time calibrated PCR assays", J Clin Microbiol, 41, pp.4565~4572 (2003)
- Bruijnesteijn Van Coppenraet ES, "Real-time PCR assay using fine-needle aspirates and tissue biopsy specimens for rapid diagnosis of mycobacterial lymphadenitis in children", J Clin Microbiol, 42, pp.2644~2650 (2004)
- Joo SI, "Evaluation of commercial PCR test kits for the detection of Mycobacterium tuberculosis from clinical specimens", J Clin Pathol Qual Control, 19, pp.237~244 (1997)
- Ozkutuk A, "Evaluation of cobas amplicor MTB test to detect Mycobacterium tuberculosis in pulmonary and extrapulmonary specimens", New Microbiol, 29, pp.269~273 (2006)
- 이기은, "항산균 배양 및 동정방법의 변화에 따른 양성률과 보고시간 변화", 대한진단검사의학회지, 25(1), pp.46~52 (2005), 대한진단검사의학회