期刊文献+

环介导等温扩增法快速检测结核分枝杆菌的初步观察 被引量:10

Application of LAMP method in rapid detection of Mycobacterium tuberculosis
在线阅读 下载PDF
导出
摘要 目的建立1种用于结核分枝杆菌快速检测的LAMP技术平台,用于结核分枝杆菌的鉴定。方法使用了11株标准菌株,建立了一套用于结核分枝杆菌快速检测的平台,对灵敏度和特异性进行优化,随后使用47株临床分离株,和13株食源性致病菌对建立的方法进行了验证。结果经过标准菌株建立并优化检测平台,经过优化的LAMP反应体系检出限为1pg的模板DNA,在使用47株临床分离株,和13株食源性致病菌对方法进行验证,发现LAMP特异性为97.4%。结论通过这项研究发现LAMP应用于结核分枝杆菌的快速检测,具有检出浓度低、特异性强和检测周期短的优点。 Objective To develop a loop-mediated isothermal amplification (LAMP) method for the detection of Mycobacterium tuberculosis. Methods 11 standard strains were used to develop the LAMP method in TB detection. The sensitivity and specificity of LAMP were evaluated using 47 M. tuberculosis and nontuberculosis mycobacteria isolates and 13 food borne strains. Results The detection limit of LAMP was 1pg of DNA,the specificity was 97.4%. Conclusions The LAMP is a rapid,sensitive,and specific method for the detection of Mycobacterium tuberculosis.
出处 《中国防痨杂志》 CAS 2010年第8期466-469,共4页 Chinese Journal of Antituberculosis
关键词 分枝杆菌 结核 核酸扩增技术 Mycobacterium tuberculosis nucleic acid amplification techniques
  • 相关文献

参考文献14

  • 1Haldar S, Chakravorty S, Bhalla M, De Majumdar S, Tyagi JS. Simplified detection of Mycobacterium tuberculosis in sputum using smear microscopy and PCR with molecular beacons[J]. J Med Mierobiol, 2007,56(10): 1356-1362.
  • 2Broceolo F, Scarpellini P, Locatelli G, Zingale A, Brambilla AM,Cichero P, Seehi LA, Malnati MS. Rapid diagnosis of mycobacterial infections and quantitation of Mycobacterium tuberculosis load by two real-time calibrated PCR assays[J]. J Clin Microbiol, 2003,41(10) :4565-4572.
  • 3Hale Y, Pfyffer G, Salfinger M. Laboratory diagnosis of mycobacterial infections: new tools and lessons learned[J]. Clin In fect Dis, 2001, 33(6): 834-846.
  • 4Pandey BD, Poudel A, Yoda T, Yoda T, Tamaru A, Oda N, Fukushima Y, Lekhak B, Risal B, Acharya B, Sapkota B, Nakajima C, Taniguchi T, Phetsuksiri B, Suzuki Y. Development of an in house loop-mediated isothermal amplification (LAMP) assay for detection of Mycobacterium tuberculosis and evaluation in sputum samples of Nepalese patients[J].J Med Microbiol, 2008, 57(4): 439-443.
  • 5Hillemann D, Warren R, Kubica T, Rtisch-Gerdes S, Niemann S. Rapid Detection of Mycobacterium tuberculosis Beijing Genotype Strains by Real-Time PCR[J]. J Clin Microbiol, 2006, 44 (9):302-306.
  • 6Shrestha NK, Tuohy MJ, Hall GS, Isada C. M. , Procop G. W.. Detection and Differentiation of Mycobacterium tuberculosis and Nontubereulous Mycobacterial Isolates by Real-Time PCR [J]. J ClinMicrobiol, 2003,41(11): 5121-5126.
  • 7Moon JW, Chang YS, Kim SK, Kim YS, Lee HM, Kim SK. Chang J. The Clinical Utility of Polymerase Chain Reaction for the Diagnosis of Pleural Tuberculosis[J]. Clin Infect Dis, 2005, 41(5), 660-666.
  • 8Notomi T, Okayama H, Masubuchi H, Yonekawa T , Watanabe K. , Amino N. , Hase T. Loop-mediated isothermal amplification of DNA[J]. Nucleic Acids Res,2000,28(12): e63.
  • 9Iwamoto T, Sonobe T, Hayashi K. Loop Mediated Isothermal Amplification for Direct Detection of Mycobacterium tuberculosis Complex, M. avium, and M. intracellulare in Sputum Samples [J]. J Clin Microbiol, 2003,41(6): 2616-2622.
  • 10Scarparo C, Piccoli P, Rigon A, Ruggiero G, Nista D, Piersimoni C. Comparison of enhanced Mycobacterium tuberculosis amplified direct test with COBAS AMPLICOR Mycobacterium tuberculosis assay for direct detection of Mycobacterium tuberculosis complex in respiratory and extrapulmonary specimens[J]. J Clin Microbiol, 2000,38(4): 1559-1562.

同被引文献95

引证文献10

二级引证文献68

相关作者

内容加载中请稍等...

相关机构

内容加载中请稍等...

相关主题

内容加载中请稍等...

浏览历史

内容加载中请稍等...
;
使用帮助 返回顶部