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焦磷酸测序技术在利福平rpoB基因突变检测中的应用评价 被引量:2

Evaluation of pyrosequencing for the detection of rpoB gene mutation in Mycobacterium tuberculosis
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摘要 目的应用焦磷酸测序技术检测结核分枝杆菌rpoB基因突变,并探讨其在结核分枝杆菌利福平耐药性检测中的应用价值。方法利用焦磷酸测序技术,测定150株结核分枝杆菌临床分离株rpoB基因81个碱基的利福平抗药性决定区(RRDR),分析rpoB基因突变特点,并与绝对浓度法和MIC法药敏结果进行比较。结果150株结核分枝杆菌中66株耐药84株敏感,54株为耐多药菌株,rpoB基因突变涉及6个密码子12种突变基因型,最常见的突变位点及突变形式分别为531位TCG突变为TTG,占60.6%,526位CAC突变为TAC、GAC,占22.7%。耐药性检测结果表明,焦磷酸检测法与绝对浓度法二者的符合率为92.7%,与MIC法的符合率为97.8%。结论焦磷酸测序检测技术不仅是一种快速、准确、高通量的利福平分子药敏检测方法,同时也是结核分枝杆菌耐药性研究的有用工具。 Objective To detect the mutations of rpoB gene in Mycobacterium tuberculosis by pyrosequencing and to evaluate the values on detection of fifampin resistance in clinical isolates. Methods Using the new technology of pyrosequencing, the mutations in the fifampin resistance determining region (RRDR) of rpoB gene were analyzed. The results were compared with those obtained from methods of the absolute concentration and the minimum inhibitory concentration(M/C). Results Among the 150 Mycobacterium tuberculosis clinical isolates,84 were susceptible and 66 resistant to RIF. 54 of the 66 resistant isolates were muhidrug-resistant (MDR) strains. Ser531Leu and His526Asp or Tyr, including twelve different genotypes and six codons,were the most common mutations. In the drug susceptibility testing, the accordance rates of the pyrosequencing and the absolute concentration method as well as MIC were 92.7% and 97.8% respectively. Conclusion Not only is the pyrosequencing technology a fast, sensitive and high throughput method in detecting rifampin resistance in Mycobacterium tuberculosis, but also a useful tool in the research of rifampin resistantce mechanism.
出处 《中华预防医学杂志》 CAS CSCD 北大核心 2008年第2期77-80,共4页 Chinese Journal of Preventive Medicine
基金 上海市科委重点专项课题(05DZ22320)
关键词 分枝杆菌 结核 微生物敏感性试验 利福平 RPOB基因 焦磷酸测序 Mycobacterium tuberculosis Microbial sensitivity test Rifampin rpoB gene Pyrosequencing
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  • 1Hopkins KL,Amold C, Threlfall EJ. Rapid detection of gyrA and parC mutations in quinolone-resistant Salmonella enterica using Pyrosequeneing technology. J Microbiol Methods, 2007, 68 : 163-171.
  • 2Haanpera M, Huovinen P,Jalava J. Detection and quantification of macrolide resistance mutations at positions 2058 and 2059 of the 23S rRNA gene by pyrosequencing. Antimicrob Agents Chemother ,2005,49 : 457-460.
  • 3中国防痨协会基础专业委员会.结核病诊断实验室检验规程.北京:中国教育出版社,2006,46-51.
  • 4郑瑞娟,秦莲花,冯永红,杨华,王洁,陆俊梅,胡忠义.焦磷酸测序技术快速检测结核分枝杆菌利福平耐药性研究[J].中华检验医学杂志,2007,30(7):737-741. 被引量:6
  • 5Garrigó M, Aragón LM, Alcaide F, et al. Multicenter laboratory evaluation of the MB/BacT Mycobacterium detection system and the BACTEC MGIT 960 system in comparison with the BACTEC 460TB system for susceptibility testing of Mycobacterium tuberculosis. J Clin Microbiol,2007 ,45 :1766-1770.
  • 6Ronaghi M, Karamohamed S, Petterssen B, et al. Real-time DNA sequencing using detection of pyrophosphate release. Anal Bioehem, 1996,242:84-89.
  • 7Ronaghi M, Uhlén M, Nyrén P. A sequencing method based on real-time pyrophosphate. Science, 1998,281 ( 5375 ) : 363,365.
  • 8刘敬华,张丽水,刘志广,黄明翔,赵秀芹,王琳,万康林.结核分枝杆菌利福平耐药基因rpoB突变特征初步分析[J].中华流行病学杂志,2006,27(11):973-976. 被引量:22
  • 9Afanas'ev MV, Ikryannikova LN, II'ina EN, et al. Molecular characteristics of rifampicin-and isoniazid-reslstant Mycobacterium tuberculosis isolates from the Russian Federation. J Antimicrob Chemother ,2007,59 : 1057-1064.
  • 10Suresh N, Singh UB, Arora J, et al. rpoB gene sequencing and spoligotyping of multidrug-resistant Mycobacterium tuberculosis isolates from India. Infect C.enet Evol,2006 ,6 :474-483 .

二级参考文献43

  • 1乐军,曾而良,谢建平,李瑶,梁莉,王洪海.中国耐多药结核分枝杆菌临床分离株rpoB基因突变特点[J].Acta Genetica Sinica,2004,31(12):1332-1336. 被引量:23
  • 2宋家武,王文栋,游风云,边壮,唐健熹,姚蓝,李本义.高通量乙型肝炎病毒YMDD突变区焦磷酸测序方法的建立及其可靠性研究[J].中华检验医学杂志,2005,28(9):957-960. 被引量:5
  • 3结核病诊断细菌学检验规程[J].中国防痨杂志,1996,18(1):28-31. 被引量:799
  • 4Espinal M A,Laszlo A,Simonsen L,Boulahbal F,Kim S J,Reniero A,Hoffner S,Rieder H L,Binkin N,Dye C,Williams R,Raviglione M C.Global trends in resistance to antituberculosis drugs.N Engl J Med,2001,344(17):1294~1303.
  • 5Dye C,Williams B G,Espinal M A,Raviglione M C.Erasing the world's slow stain: strategies to beat multidrug-resistant tuberculosis.Science,2002,295(5562):2042~2046.
  • 6Becerra M C,Bayona J,Freeman J,Farmer P E,Kim J Y.Redefining MDR-TB transmission-drug resistance'hot spots'.Int J Tuberc Lung Dis,2000,4(5): 387~394.
  • 7Dye C,Scheele S,Dolin P,Pathania V,Raviglione M C.Consensus statement.Global burden of tuberculosis: estimated incidence,prevalence,and mortality by country.WHO Global Surveillance and Monitoring Project.JAMA,1999,18:677~686.
  • 8Dye C,Williams B G.Criteria for the control of drug-resistant tuberculosis.Proc Natl Acad Sci USA,2000,97(14):8180~8185.
  • 9Drobniewski F A,Wilson S M.The rapid diagnosis of isoniazid and rifampin resistance in Mycobacterium tuberculosis--a molecular story.J Med Microbiol,1998,47(3): 189~196.
  • 10Ramaswamy S,Musser J M.Molecular genetic basis of antimicrobial agent resistance in Mycobacterium tuberculosis:1998 update.Tuberc Lung Dis,1998,79:3~29.

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