Denaturing HPLC를 이용한 리팜핀 내성 결핵균의 검출
Detection of Rifampin Resistant Mycobacterium tuberculosis complex using Denaturing HPLC
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Abstract
Background : Tuberculosis (TB) remains an important cause of morbidity and mortality throughout the world. The surge of TB has been accompanied by an increase in multi-drug-resistant tuberculosis (MDR-TB). In this study, we developed a denaturing HPLC (DHPLC) method for detecting rpoB gene mutation as a rifampin resistance based on sequence. Methods : In this study, we used 99 mycobacterial isolates grown in Ogawa media. At first, we used a PCR method that can amplify the 235 bp and 136 bp rpoB DNAs of Mycobacterium tuberculosis complex (MTB) and Non-tuberculous mycobacteria (NTM). And then, PCR-restriction fragment length polymorphism (RFLP) of rpoB DNA (342 bp), which comprises the RifT region, was used for the differential identification of Mycobacteria. Finally, we detected these amplicons by DHPLC, compared to PCR-RFLP results, and performed sequencing. Results : Among 99 mycobacterial isolates, 80 (81%) were MTB and 19 (19%) were NTM. NTM were identified to 7 different species by DHPLC and PCR-RFLP. rpoB mutation was detected in 9 (11%) of the MTB specimens. These results were confirmed by using sequencing. Conclusions : DHPLC provided a rapid, simple, and automatable performance for detection of rifampin resistant Mycobacterium tuberculosis complex and would be helpful as a supplemental method in high-throughput clinical laboratories. (Korean J Lab Med 2008;28:95-102)
참고문헌 (30)
- Nunn P, "Tuberculosis and HIV infection: the global setting", J Infect Dis, 196(S), pp.S5~14 (2007)
- Korea National Statistical Office, "Annual report on the cause of death statistics" (2003), Based on vital registration
- Barnes PF, "Tuberculosis in patients with human immuno-deficiency virus infection", N Engl J Med, 324, pp.1644~1650 (1991)
- Cantwell MF, "Onorato IM. Epidemiology of tuberculosis in the United States, 1985 through 1992", JAMA, 272, pp.535~539 (1994)
- Starke JR, "Resurgence of tuberculosis in children", J Pediatr, 120, pp.839~855 (1992)
- Morris S, "Molecular mechanisms of multiple drug resistance in clinical isolates of Mycobacterium tuberculosis", J Infect Dis, 171, pp.954~960 (1995)
- Espinal MA, "Global trends in resistance to antituberculosis drugs. World Health Organization-International Union against Tuberculosis and Lung Disease Working Group on Anti-Tuberculosis Drug Resis tance Surveillance", N Engl J Med, 344, pp.1294~1303 (2001)
- Hunt JM, "Detection of a genetic locus encoding resistance to rifampin in mycobacterial cultures and in clinical specimens", Diagn Microbiol Infect Dis, 18, pp.219~227 (1994)
- Donnabella V, "Isolation of the gene for the beta subunit of RNA polymerase from rifampicin-resistant Mycobacterium tuberculosis and identification of new mutations", Am J Respir Cell Mol Biol, 11, pp.639~643 (1994)
- Goble M, "Treatment of 171 patients with pulmonary tuberculosis resistant to isoniazid and rifampin", N Engl J Med, 328, pp.527~532 (1993)
- Lee MK, "Rapid Detection of rifampin Resistant Mycobacterium tuberculosis using the Lind probe Assay", Korean J Clin Pathol, 17, pp.269~278 (1997)
- Underhill PA, "A pre- Columbian Y chromosome-specific transition and its implications for human evolutionary history", Proc Natl Acad Sci USA, 93, pp.196~200 (1996)
- 김종규, "DHPLC의 화학적 특성을 이용한 질병 유전자의 분석 시스템 개발", 대한화학회지, 50(2), pp.116~122 (2006), 대한화학회
- Kim BJ, "Differentiation of mycobacterial species by PCR-restriction analysis of DNA (342 base pairs) of the RNA polymerase gene (rpoB)", J Clin Microbiol, 39, pp.2102~2109 (2001)
- Kim BJ, "Differential identification of Mycobacterium tuberculosis complex and nontuberculous mycobacteria by duplex PCR assay using the RNA polymerase gene (rpoB)", J Clin Microbiol, 42, pp.1308~1312 (2004)
- Kam KM, "Surveillance of Mycobacterium tuberculosis drug resistance in Hong Kong, 1986-1999, after the implementation of directly observed treatment", Int J Tuberc Lung Dis, 5, pp.815~823 (2001)
- Xiao W, "Denaturing high-performance liquid chromatography: A review", Hum Mutat, 17, pp.439~474 (2001)
- Skopek TR, "Analysis of sequence alterations in a defined DNA region: comparison of temperature- modulated heteroduplex analysis and denaturing gradient gel electrophoresis", Mutat Res, 430, pp.13~21 (1999)
- Jones AC, "Optimal temperature selection for mutation detection by denaturing HPLC and comparison to single-stranded conformation polymorphism and heteroduplex analysis", Clin Chem, 45, pp.1133~1140 (1999)
- Yip CW, "Denaturing HPLC for high-throughput screening of rifampicinresistant Mycobacterium tuberculosis isolates", Int J Tuberc Lung Dis, 10, pp.625~630 (2006)
- Wagner T, "Denaturing high-performance liquid chromatography detects reliably BRCA1 and BRCA2 mutations", Genomics, 62, pp.369~376 (1999)
- Ellis LA, "A comparison of fluorescent SSCP and denaturing HPLC for high throughput mutation scanning", Hum Mutat, 15, pp.556~564 (2000)
- Gross E, "Identification of specific BRCA1 and BRCA2 variants by DHPLC", Hum Mutat, 16, pp.345~353 (2000)
- Nickerson ML, "Signature-based analysis of MET proto-oncogene mutations using DHPLC", Hum Mutat, 16, pp.68~76 (2000)
- Benit P, "Mutation analysis of the hamartin gene using denaturing high performance liquid chromatography", Hum Mutat, 16, pp.417~421 (2000)
- 이정연, "한 대학병원에서 비결핵항산균의 분리 및 동정실태", 결핵 및 호흡기질환, 58(4), pp.385~391 (2005), 대한결핵및호흡기학회
- Yip CW, "Denaturing HPLC for high-throughput screening of rifampicinresistant Mycobacterium tuberculosis isolates", Int J Tuberc Lung Dis, 10, pp.625~630 (2006)
- Telenti A, "Detection of rifampicin-resistance mutations in Mycobacterium tuberculosis", Lancet, 341, pp.647~650 (1993)
- Suh JT, "Direct sequencing for rpoB gene in rifampin-resistant Mycobacterium tuberculosis", Korean J Clin Pathol, 16, pp.78~90 (1996)
- Lee MA, "Direct detection of rifampin-resistant mycobacterium tuberculosis by polymerase chain reaction and line probe assay in sputums", Korean J Clin Pathol, 18, pp.71~76 (1998)