RNA恒温扩增实时荧光检测技术对复治菌阳肺结核患者疗效评价的研究
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摘要
目的探讨RNA恒温扩增实时荧光检测(SAT)技术在初次复治菌阳肺结核患者化疗效果监测中的价值。方法选取2015年6月—2016年12月在我科住院,临床诊断为初次复治菌阳肺结核患者62例。患者经标准化的抗结核治疗2个月后,取痰标本分别经彭氏杯抗酸染色、BACTEC MGIT-960培养和菌种鉴定、SAT检测,以BACTECMGIT-960培养、菌种鉴定为金标准,评价SAT在抗结核治疗药物疗效监测方面的价值。结果 62例患者治疗后结核菌培养阳性49例,其中人型结核分枝杆菌42例,非结核分枝杆菌7例;结核菌培养阴性13例。痰彭氏杯抗酸染色阳性32例,阴性30例。SAT检测阳性41例,阴性21例。SAT与痰培养有较好的一致性(Kappa值=0.964),其敏感度、特异度、阳性预测值、阴性预测值分别为97.62%、100%、100%、95.24%;彭氏杯抗酸染色与MGIT-960培养结果一致性较差(Kappa值=0.086),其敏感度、特异度、阳性预测值、阴性预测值分别为54.76%、55.00%、71.88%、36.67%。结论SAT技术可用于初次复治菌阳肺结核患者药物疗效的监测,可根据其结果及时调整用药,值得临床推广。
Objective To investigate the value of real-time fluorescence detection technique of RNA(SAT) constant amplification in monitoring the effect of chemotherapy on patients with sputum positive pulmonary tuberculosis.Methods Sixty-two patients were selected, who were clinically diagnosed as the first-time retreatment for sputum smear-positive pulmonary tuberculosis and were hospitalized in our department from June 2015 to December 2016. After two-monthst andard anti-tuberculosis treatment, sputum samples were detected by Peng's vessel acid-fast staining, BACTEC MGIT-960 culture and strain identification, SAT detection. The BACTEC MGIT-960 culture and strain identification were used as gold standards, the value of SAT during the monitoring the therapeutic effect of the anti-tuberculosis drugs was assessed.Results After the treatment, 49 cases out of 62 showed positive results in mycobacterium tuberculosis culture test, among them 42 patients were diagnosed as human type mycobacterium tuberculosis, 7 patients were diagnosed as nontuberculous mycobacteria infection, and 13 cases showed negative results in mycobacterium tuberculosis culture. Thirty-two cases showed positive results in sputum Peng's vessel acid-fast staining test, and 30 cases showed negative results. Forty-one cases showed positive results in SAT test and 21 cases showed negative results in SAT. SAT results were well concordant with sputum culture results(Kappa value=0.964), and the sensitivity, the specificity, the positive predictive value and thenegative predictive value of SAT were 97.62%, 100%, 100% and 95.24% respectively. Peng's vessel acid-fast stainingresults were badly concordant with MGIT-960 culture results(Kappa value=0.086), and the sensitivity, the specificity, the positive predictive value and the negative predictive value of Peng's vessel acid-fast staining were 54.76%, 55.00%,71.88% and 36.67% respectively.Conclusion SAT results can be used as good indices during the monitoring therapeutic effects of drugs used for the first-time retreatment in patients with sputum smear-positive pulmonary tuberculosis, which is worth promoting.
Objective To investigate the value of real-time fluorescence detection technique of RNA(SAT) constant amplification in monitoring the effect of chemotherapy on patients with sputum positive pulmonary tuberculosis.Methods Sixty-two patients were selected, who were clinically diagnosed as the first-time retreatment for sputum smear-positive pulmonary tuberculosis and were hospitalized in our department from June 2015 to December 2016. After two-monthst andard anti-tuberculosis treatment, sputum samples were detected by Peng's vessel acid-fast staining, BACTEC MGIT-960 culture and strain identification, SAT detection. The BACTEC MGIT-960 culture and strain identification were used as gold standards, the value of SAT during the monitoring the therapeutic effect of the anti-tuberculosis drugs was assessed.Results After the treatment, 49 cases out of 62 showed positive results in mycobacterium tuberculosis culture test, among them 42 patients were diagnosed as human type mycobacterium tuberculosis, 7 patients were diagnosed as nontuberculous mycobacteria infection, and 13 cases showed negative results in mycobacterium tuberculosis culture. Thirty-two cases showed positive results in sputum Peng's vessel acid-fast staining test, and 30 cases showed negative results. Forty-one cases showed positive results in SAT test and 21 cases showed negative results in SAT. SAT results were well concordant with sputum culture results(Kappa value=0.964), and the sensitivity, the specificity, the positive predictive value and thenegative predictive value of SAT were 97.62%, 100%, 100% and 95.24% respectively. Peng's vessel acid-fast stainingresults were badly concordant with MGIT-960 culture results(Kappa value=0.086), and the sensitivity, the specificity, the positive predictive value and the negative predictive value of Peng's vessel acid-fast staining were 54.76%, 55.00%,71.88% and 36.67% respectively.Conclusion SAT results can be used as good indices during the monitoring therapeutic effects of drugs used for the first-time retreatment in patients with sputum smear-positive pulmonary tuberculosis, which is worth promoting.
引文
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[6]孙宝彬,谢明娟,陈效友.恒温扩增实时荧光检测技术检测痰液对肺结核患者诊断价值的Meta分析[J].中国防痨杂志,2017,39(5):511-517.Sun BB,Xie MJ,Chen XY.The diagnostic value of SAT-TB for detection sputum in patients with pulmonary tuberculosis:a Meta-analysis[J].Chinese Journal of Antituberculosis,2017,39(5):511-517.doi:10.3969/j.issn.1000-6621.2017.05.019.
[7]中华医学会.临床诊疗指南(结核病分册)[M].北京:人民卫生出版社,2004:80.Chinese Medical Association.Guidelines for clinical diagnosis and treatment(tuberculosis register)[M].Beijing:People’s Health Publishing House,2004:80.
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[10]张慧慧,熊国亮.RNA恒温扩增实时荧光检测技术检测肺结核患者痰标本中结核分枝杆菌的临床应用研究[J].实验与检验医学,2015,33(4):407-409.Zhang HH,Xiong GL.Evaluation of the isothermal RNA amplification assay for detection of Mycobacterium tuberculosis in sputum specimens[J].Experimental and Laboratory Medicine,2015,33(4):407-409.doi:10.3969/j.issn.1674-1129.2015.04.007.
[11]沙巍,何娅,蒋瑞华,等.实时荧光核酸恒温放大检测(SAT)法对肺结核诊断价值的研究[J].中国防痨杂志,2012,34(6):377-379.Sha W,He Y,Jiang RH,et al.Diagnostic value of simultaneous amplification and testing in patients with pulmonary tuberculosis[J].Chinese Journal of Antituberculosis,2012,34(6):377-379.
[12]中国防痨协会临床专业委员会.结核病临床诊治进展报告(年)(第一部分结核病临床诊断)[]中国防痨杂志,2013,35(6):405-426.Chinese Anti-tuberculosis Association of Clinical Society.Annual report on clinical diagnosis and treatment progress of tuberculosis(2012)(Part 1 clinical diagnosis)[J].Chinese Journal of Antituberculosis,2013,35(6):405-426.
[13]Pollock N,Westerling J,Sloutsky A.Specimen dilution increases the diagnostic utility of the gen-probe mycobacterium tuberculosis direct test[J].Am J Clin Pathol,2006,126:142-147.doi:10.1309/JBQH-WC6H-4YN6-F67Q.
[14]Piersimoni C,Scarparo C,Piccoli P,et al.Performance assessment of two commercial amplification assays for direct detection of Mycobacterium tuberculosis complex from respiratory and extrapulmonary specimens[J].J Clin Microbiol,2002,40:4138-4142.doi:10.1128/JCM.40.11.4138-4142.2002.
[15]Cui Z,Wang Y,Fang L,et al.Novel real-time simultaneous amplification and testing method to accurately and rapidly detect Mycobacterium tuberculosis complex[J].J Clin Microbiol,2012,50(3):646-650.doi:10.1128/JCM.05853-11.
[16]胡永领,刘成永,成松.应用RNA恒温扩增实时荧光检测技术观察抗结核药物的疗效[J].海峡药学,2015,27(1):75-77.Hu YL,Liu CY,Cheng S.Application of isothermal RNA amplification real-time fluorescence detection technology to observe the curative effect of anti-TB drugs[J].Strait Pharmaceutical Journal,2015,27(1):75-77.doi:10.3969/j.issn.1006-3765.2015.01.032.
[17]王永忠,张宏宇,刘小琴,等.痰液标本结核分支杆菌RNA恒温扩增检测及临床应用[J].山东医药,2011,51(41):83-84.Wang YZ,Zhang HY,Liu XQ,et al.Detection of Mycobacterium tuberculosis RNA in sputum by isothermal amplification and its clinical application[J].Shandong Medical Journal,2011,51(41):83-84.doi:10.3969/j.issn.1002-266X.2011.41.048.
[2]陈婷婷,钱煦岱,包文婷,等.SAT技术检测解脲脲原体和沙眼衣原体核酸在泌尿生殖道感染中的应用[J].中国卫生检验杂志,2015,25(14):2357-2359.Chen TT,Qian XD,Bao WT,et al.Application of SAT technology in the detection of Ureaplasma urealyticum and Chlamydia trachomatis nulcleic acid in patients with urogenital tract infection[J].Chinese Journal of Health Laboratory Technology,2015,25(14):2357-2359.
[3]朱海燕,冯金梦,冯美卿,等.实时荧光核酸恒温检测技术(SAT)在甲型流感病毒动物模型检测中的应用[J].工业微生物,2014,44(2):63-68.Zhu HY,Feng JM,Feng MQ,et al.Application of simultaneous amplification and testing(SAT)in detection of influenza A experimental animal model[J].Industrial Microbiology,2014,44(2):63-68.doi:10.3969/j.issn.1001-6678.2014.02.012.
[4]曹凌峰,苏犁云,董妞妞,等.荧光恒温扩增技术检测肠道病毒71型方法的建立与评价[J].检验医学,2014,29(11):1115-1119.Cao LF,Su LY,Dong NN,et al.Establishment and evaluation of fluorescence isothermal amplification assay for the detection of Enterovirus 71[J].Laboratory Medicine,2014,29(11):1115-1119.doi:10.3969/j.issn.1673-8640.2014.11.009.
[5]高志华,金印,陈峰.实时荧光核酸恒温扩增技术检测尿液中淋球菌的分析[J].国际检验医学杂志,2012,33(4):463-464.Gao ZH,Jin Y,Chen F.Detection of Neisseria gonorrhoeae in urine by real-time fluorescence nucleic acid isothermal amplification[J].International Journal of Laboratory Medicine,2012,33(4):463-464.doi:10.3969/j.issn.1673-4130.2012.04.039.
[6]孙宝彬,谢明娟,陈效友.恒温扩增实时荧光检测技术检测痰液对肺结核患者诊断价值的Meta分析[J].中国防痨杂志,2017,39(5):511-517.Sun BB,Xie MJ,Chen XY.The diagnostic value of SAT-TB for detection sputum in patients with pulmonary tuberculosis:a Meta-analysis[J].Chinese Journal of Antituberculosis,2017,39(5):511-517.doi:10.3969/j.issn.1000-6621.2017.05.019.
[7]中华医学会.临床诊疗指南(结核病分册)[M].北京:人民卫生出版社,2004:80.Chinese Medical Association.Guidelines for clinical diagnosis and treatment(tuberculosis register)[M].Beijing:People’s Health Publishing House,2004:80.
[8]张贺秋,赵雁林.现代结核病诊断技术[M].北京:人民卫生出版社,2013:282-285.Zhang HQ,Zhao YL.Modern diagnostic techniques for tuberculosis[M].Beijing:People’s Health Publishing House,2013:282-285.
[9]World Health Organization.Global tuberculosis report 2016[EB/OL].Geneva:World Health Organization,2016.http://apps.who.int/iris/bitstream/handle/10665/250441/9789241565394-eng.pdf;jsessionid=15FB31E063F2265EB20F8845589C4DC7?sequence=1.
[10]张慧慧,熊国亮.RNA恒温扩增实时荧光检测技术检测肺结核患者痰标本中结核分枝杆菌的临床应用研究[J].实验与检验医学,2015,33(4):407-409.Zhang HH,Xiong GL.Evaluation of the isothermal RNA amplification assay for detection of Mycobacterium tuberculosis in sputum specimens[J].Experimental and Laboratory Medicine,2015,33(4):407-409.doi:10.3969/j.issn.1674-1129.2015.04.007.
[11]沙巍,何娅,蒋瑞华,等.实时荧光核酸恒温放大检测(SAT)法对肺结核诊断价值的研究[J].中国防痨杂志,2012,34(6):377-379.Sha W,He Y,Jiang RH,et al.Diagnostic value of simultaneous amplification and testing in patients with pulmonary tuberculosis[J].Chinese Journal of Antituberculosis,2012,34(6):377-379.
[12]中国防痨协会临床专业委员会.结核病临床诊治进展报告(年)(第一部分结核病临床诊断)[]中国防痨杂志,2013,35(6):405-426.Chinese Anti-tuberculosis Association of Clinical Society.Annual report on clinical diagnosis and treatment progress of tuberculosis(2012)(Part 1 clinical diagnosis)[J].Chinese Journal of Antituberculosis,2013,35(6):405-426.
[13]Pollock N,Westerling J,Sloutsky A.Specimen dilution increases the diagnostic utility of the gen-probe mycobacterium tuberculosis direct test[J].Am J Clin Pathol,2006,126:142-147.doi:10.1309/JBQH-WC6H-4YN6-F67Q.
[14]Piersimoni C,Scarparo C,Piccoli P,et al.Performance assessment of two commercial amplification assays for direct detection of Mycobacterium tuberculosis complex from respiratory and extrapulmonary specimens[J].J Clin Microbiol,2002,40:4138-4142.doi:10.1128/JCM.40.11.4138-4142.2002.
[15]Cui Z,Wang Y,Fang L,et al.Novel real-time simultaneous amplification and testing method to accurately and rapidly detect Mycobacterium tuberculosis complex[J].J Clin Microbiol,2012,50(3):646-650.doi:10.1128/JCM.05853-11.
[16]胡永领,刘成永,成松.应用RNA恒温扩增实时荧光检测技术观察抗结核药物的疗效[J].海峡药学,2015,27(1):75-77.Hu YL,Liu CY,Cheng S.Application of isothermal RNA amplification real-time fluorescence detection technology to observe the curative effect of anti-TB drugs[J].Strait Pharmaceutical Journal,2015,27(1):75-77.doi:10.3969/j.issn.1006-3765.2015.01.032.
[17]王永忠,张宏宇,刘小琴,等.痰液标本结核分支杆菌RNA恒温扩增检测及临床应用[J].山东医药,2011,51(41):83-84.Wang YZ,Zhang HY,Liu XQ,et al.Detection of Mycobacterium tuberculosis RNA in sputum by isothermal amplification and its clinical application[J].Shandong Medical Journal,2011,51(41):83-84.doi:10.3969/j.issn.1002-266X.2011.41.048.