PCR-Single-Strand Conformational Polymorphism Method for Detecting rpoB Gene of Rifampin-Resistant Mycobacterium Tuberculosis: A Meta-Analysis_Chinese Journal of Evidence-Based Medicine

Authors：

 YULu , MENGCun-ren ,  ZHANGZhao-xia

Department of Laboratory Medicine, First Affiliated Hospital of Xinjiang Medical University, Urumqi 830054, China;

Corresponding author：

ZHANGZhao-xia, Email: 285715300@qq.com

Keywords：

Mycobacterium tuberculosis; Rifampin; rpoB; Polymerase chain reaction-single strand conformation polymorphism; Meta-analysis

DOI：

10.7507/1672-2531.20140119

Video：

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Objective To systematically review the diagnostic value of PCR-single-strand conformational polymorphism (PCR-SSCP) method for detecting rpoB gene mutation of rifampin-resistant mycobacterium tuberculosis. Methods Such databases as PubMed, Web of Science, The Cochrane Library (Issue 2, 2014), CBM, VIP and WanFang Data were electronically and comprehensively searched for relevant studies on the diagnostic value of PCR-SSCP method for detecting rpoB gene mutation of rifampin-resistant mycobacterium tuberculosis from inception to January 1^st, 2014. Literature screening according to the inclusion and exclusion criteria, data extraction and methodological quality assessment were completed by two reviewers independently. Meta-analysis was then conducted using Meta-Disc 1.4 and Stata 12.0. Results A total of 10 studies were included involving 1 299 cases. The results of meta-analysis showed SEN=0.92 (95%CI 0.90 to 0.94, P=0.019 3), SPE=0.97 (95%CI 0.95 to 0.98, P < 0.000 1), +LR=23.68 (95%CI 8.71 to 64.37, P < 0.000 1), -LR=0.10 (95%CI 0.06 to 0.15, P=0.023 1), DOR=257.16 (95%CI 96.82 to 683.02, P=0.020 0), and SROC area under the curve (AUC) was 0.971 5, and Q* was 0.922 3. The results of sensitivity analysis (after removing studies with sample size less than 100, Chinese studies and QUADAS more than 10 studies) showed that, the results were stable with reliable conclusion. Conclusion PCR-SSCP method has a fairly high value in the diagnosis of rpoB gene mutation of rifampinresistant mycobacterium tuberculosis.

Citation： YULu, MENGCun-ren, ZHANGZhao-xia. PCR-Single-Strand Conformational Polymorphism Method for Detecting rpoB Gene of Rifampin-Resistant Mycobacterium Tuberculosis: A Meta-Analysis. Chinese Journal of Evidence-Based Medicine, 2014, 14(6): 698-706. doi: 10.7507/1672-2531.20140119 Copy

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2.	孟繁荣, 刘志辉, 谢贝, 等.我国利福平敏感结核杆菌rpoB基因利福平耐药定区序列循证分析.实用医学杂志, 2009, 29(21):3601-3604.
3.	Miller LP, Crawford JT, Shinnick TM. The rpoB gene of Mycobacterium tuberculosis. Ntimicrob Agents Ch, 1994, 38(4):805-811.
4.	Palomino J C. Nonconventional and new methods in the diagnosis of tuberculosis:feasibility and applicability in the field. Eur Respir J, 2005, 26(2):339-350.
5.	Palomino JC. Newer diagnostics for tuberculosis and multi-drug resistant tuberculosis. Curr Opin Pulm Med, 2006, 12(3):172-178.
6.	Isfahani BN, Tavakoli A, Salehi M, et al. Detection of rifampin resistance patterns in Mycobacterium tuberculosis strains isolated in Iran by polymerase chain reaction-single-strand conformation polymorphism and direct sequencing methods. Mem Inst Oswaldo Cruz, Rio de Janeiro, 2006, 101(6):597-602.
7.	Scarpellini P, Braglia S, Brambilla AM, et al. Detection of rifampin resistance by single-strand conformation polymorphism analysis of cerebrospinal fluid of patients with tuberculosis of the central nervous system. J Clin Microbiol, 1997, 35(11):2802-2806.
8.	汪保国, 陈思东, 张胜, 等.结核分支杆菌rpoB基因突变与对利福平耐药的关系.实用医学杂志, 2009, 25(7):1022-1024.
9.	Whiting P, Rutjes AW, Reitsma IB, 等. QUADAS的制定:用于系统评价中评价诊断性研究质量的工具(译文).中国循证医学杂志, 2007, 7(4):296-306.
10.	Whiting P, Weswood ME, Rutjes AW, 等. QUADAS评价:一种用于诊断性研究的质量评价工具(修订版)(译文).中国循证医学杂志, 2007, 7(7):531-536.
11.	张天嵩, 钟文昭. Meta Disc软件在诊断试验Meta分析中的应用.循证医学, 2008, 8(2):97-108.
12.	杨柳, 苏明权, 程晓东, 等. PCR-SSCP法检测结核分枝杆菌耐药性.第四军医大学学报, 2004, 24(23):2142-214.
13.	钟敏, 温博海.检测结核分枝杆菌rpoB基因突变的研究.中华检验医学杂志, 2002, 25(1):38-41.
14.	褚美芬, 严杰, 钟雅文, 等.快速检测结核分枝杆菌异烟肼和利福平耐药相关基因mPCR-SSCP建立及应用.中华微生物学和免疫学杂志, 2012, 32(7):655-660.
15.	Cheng X, Zhang J, Yang L, et al. A new Multi-PCR-SSCP assay for simultaneous detection of isoniazid and rifampin resistance in mycobacterium tuberculosis. J Microbiol Meth, 2007, 70:301-305.
16.	Telenti A, Honore N, Bernasconi C, et al. Genotypic assessment of isoniazid and rifampin resistance in mycobacterium tuberculosis: a blind study at reference laboratory level. J Clin Microbiol, 1997, 35(3):719-723.
17.	Sheng J, Li J, Sheng G, et al. Characterization of rpoB mutations associated with rifampin resistance in Mycobacterium tuberculosis from eastern China. J Appl Microbiol, 2008, 105(3):904-911.
18.	Negi SS, Rai A, Gupta S, et al. Characterization of rpoB gene for detection of rifampicin drug resistance by SSCP and sequence analysis. Int J Infect Dis, 2008, 12:e318.
19.	Mani C, Selvakumar N, Kumar V, et al. Comparison of DNA sequencing, PCR-SSCP and PhaB assays with indirect sensitivity testing for detection of rifampicin resistance in mycobacterium tuberculosis. Int J Tuberc Lung D, 2003, 7(7):652-659.
20.	Kim BJ, Lee KH, Yun YJ, et al. Simultaneous identification of rifampin-resistant Mycobacterium tuberculosis and nontuberculous mycobacteria by polymerase chain reaction-single strand conformation polymorphism and sequence analysis of the RNA polymerase gene (rpoB). J Microbiol Meth, 2004, 58(1):111-118.
21.	Wu X, Zhang J, Zhuang Y, et al. Molecular mechanisms of drug resistance in mycobacterium tuberculosis clinical isolates. CMJ, 1999, 112(6):524-528.
22.	Devillé WL, Buntinx F, Bouter LM, et al. Conducting systematic reviews of diagnostic studies:didactic guidelines. BMC Medical Research Methodology, 2002, 2(1):1-13.
23.	柳清云, 孙刚, 高谦.结核分枝杆菌(MTB)异质性耐药研究进展.复旦学报(医学版), 2013, 40(1):1-4.
24.	Bossuyt PMM, Reitsma JB, Bruns DE, et al. The STARD statement for reporting studies of diagnostic accuracy:Explanation and elaboration. Clin Chem, 2003, 49(1):7-18.
25.	Tackmann R'Schuetz G, HammB, et al. Quality of the reporting of diagnostic accuracy studies:STARD (Standards for the Reporting of Diagnostic accuracy studies). Rofo, 2010, 182(8):655-659.

1. Zumla A, Abubakar I, Raviglione M, et al. Drug-resistant tuberculosis-current dilemmas, unanswered questions, challenges, and priority needs. J Infect Dis, 2012, 205(2):228-240.
2. 孟繁荣, 刘志辉, 谢贝, 等.我国利福平敏感结核杆菌rpoB基因利福平耐药定区序列循证分析.实用医学杂志, 2009, 29(21):3601-3604.
3. Miller LP, Crawford JT, Shinnick TM. The rpoB gene of Mycobacterium tuberculosis. Ntimicrob Agents Ch, 1994, 38(4):805-811.
4. Palomino J C. Nonconventional and new methods in the diagnosis of tuberculosis:feasibility and applicability in the field. Eur Respir J, 2005, 26(2):339-350.
5. Palomino JC. Newer diagnostics for tuberculosis and multi-drug resistant tuberculosis. Curr Opin Pulm Med, 2006, 12(3):172-178.
6. Isfahani BN, Tavakoli A, Salehi M, et al. Detection of rifampin resistance patterns in Mycobacterium tuberculosis strains isolated in Iran by polymerase chain reaction-single-strand conformation polymorphism and direct sequencing methods. Mem Inst Oswaldo Cruz, Rio de Janeiro, 2006, 101(6):597-602.
7. Scarpellini P, Braglia S, Brambilla AM, et al. Detection of rifampin resistance by single-strand conformation polymorphism analysis of cerebrospinal fluid of patients with tuberculosis of the central nervous system. J Clin Microbiol, 1997, 35(11):2802-2806.
8. 汪保国, 陈思东, 张胜, 等.结核分支杆菌rpoB基因突变与对利福平耐药的关系.实用医学杂志, 2009, 25(7):1022-1024.
9. Whiting P, Rutjes AW, Reitsma IB, 等. QUADAS的制定:用于系统评价中评价诊断性研究质量的工具(译文).中国循证医学杂志, 2007, 7(4):296-306.
10. Whiting P, Weswood ME, Rutjes AW, 等. QUADAS评价:一种用于诊断性研究的质量评价工具(修订版)(译文).中国循证医学杂志, 2007, 7(7):531-536.
11. 张天嵩, 钟文昭. Meta Disc软件在诊断试验Meta分析中的应用.循证医学, 2008, 8(2):97-108.
12. 杨柳, 苏明权, 程晓东, 等. PCR-SSCP法检测结核分枝杆菌耐药性.第四军医大学学报, 2004, 24(23):2142-214.
13. 钟敏, 温博海.检测结核分枝杆菌rpoB基因突变的研究.中华检验医学杂志, 2002, 25(1):38-41.
14. 褚美芬, 严杰, 钟雅文, 等.快速检测结核分枝杆菌异烟肼和利福平耐药相关基因mPCR-SSCP建立及应用.中华微生物学和免疫学杂志, 2012, 32(7):655-660.
15. Cheng X, Zhang J, Yang L, et al. A new Multi-PCR-SSCP assay for simultaneous detection of isoniazid and rifampin resistance in mycobacterium tuberculosis. J Microbiol Meth, 2007, 70:301-305.
16. Telenti A, Honore N, Bernasconi C, et al. Genotypic assessment of isoniazid and rifampin resistance in mycobacterium tuberculosis: a blind study at reference laboratory level. J Clin Microbiol, 1997, 35(3):719-723.
17. Sheng J, Li J, Sheng G, et al. Characterization of rpoB mutations associated with rifampin resistance in Mycobacterium tuberculosis from eastern China. J Appl Microbiol, 2008, 105(3):904-911.
18. Negi SS, Rai A, Gupta S, et al. Characterization of rpoB gene for detection of rifampicin drug resistance by SSCP and sequence analysis. Int J Infect Dis, 2008, 12:e318.
19. Mani C, Selvakumar N, Kumar V, et al. Comparison of DNA sequencing, PCR-SSCP and PhaB assays with indirect sensitivity testing for detection of rifampicin resistance in mycobacterium tuberculosis. Int J Tuberc Lung D, 2003, 7(7):652-659.
20. Kim BJ, Lee KH, Yun YJ, et al. Simultaneous identification of rifampin-resistant Mycobacterium tuberculosis and nontuberculous mycobacteria by polymerase chain reaction-single strand conformation polymorphism and sequence analysis of the RNA polymerase gene (rpoB). J Microbiol Meth, 2004, 58(1):111-118.
21. Wu X, Zhang J, Zhuang Y, et al. Molecular mechanisms of drug resistance in mycobacterium tuberculosis clinical isolates. CMJ, 1999, 112(6):524-528.
22. Devillé WL, Buntinx F, Bouter LM, et al. Conducting systematic reviews of diagnostic studies:didactic guidelines. BMC Medical Research Methodology, 2002, 2(1):1-13.
23. 柳清云, 孙刚, 高谦.结核分枝杆菌(MTB)异质性耐药研究进展.复旦学报(医学版), 2013, 40(1):1-4.
24. Bossuyt PMM, Reitsma JB, Bruns DE, et al. The STARD statement for reporting studies of diagnostic accuracy:Explanation and elaboration. Clin Chem, 2003, 49(1):7-18.
25. Tackmann R'Schuetz G, HammB, et al. Quality of the reporting of diagnostic accuracy studies:STARD (Standards for the Reporting of Diagnostic accuracy studies). Rofo, 2010, 182(8):655-659.

Chinese Journal of Evidence-Based Medicine

PCR-Single-Strand Conformational Polymorphism Method for Detecting rpoB Gene of Rifampin-Resistant Mycobacterium Tuberculosis: A Meta-Analysis

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