GenoType MTBDRplus方法对利福平耐药检测研究

发布时间：2019-06-03 09:09

【摘要】：目的：应用GenoType MTBDRplus方法和传统比例法,对结核分枝杆菌利福平耐药性进行检测研究对比。GenoType MTBDRplus方法采用核酸反向线性探针杂交技术检测与利福平耐药密切相关的rpoB基因,可在24小时内完成检测。传统比例法需要3个月左右完成检测,虽然方法简单、经济、易于推广,难于满足临床对结核病诊断和治疗的需求,限制了其临床的应用。GenoType MTBDRplus方法为早期诊断耐药结核病提供快速检测依据,利于临床早期合理治疗。 方法：标本来自于2011年4月至2011年9月,天津市滨海新区结核病防治所就诊涂片阳性患者的痰标本,共收集55例患者的痰标本。同一份痰标本分别进行GenoType MTBDRplus方法和传统比例法检测,检测结核分枝杆菌对利福平耐药性。GenoType MTBDRplus方法采用核酸反向线性探针杂交技术检测与利福平耐药密切相关的rpoB基因,推测其对利福平的耐药性。GenoType MTBDRplus方法检测过程包括,涂阳痰标本的处理通过碱处理-中和离心沉淀法,使用GenoLyse试剂盒提取DNA,PCR扩增,杂交,结果判读完成。传统比例法检测结核分枝杆菌利福平耐药性,通过实验材料准备,制备菌悬液和稀释,接种菌液和培养,结果报告完成。应用Kappa检验对两种方法进行一致性的统计分析。研究在天津市结核病控制中心参比室进行。 结果：1.55例涂阳标本中,1例经鉴定为非结核分枝杆菌(NTM),1例培养结果为污染,1例培养阳性而GenoType MTBDRplus无法判读。可与比例法药物敏感性试验进行对比的标本为52例。2.利福平耐药性的结果。以比例法药敏结果为判断标准,使用GenoType MTBDRpuls检测利福平耐药性的敏感度为75%,特异度为97.9%,阳性预测值为75%,阴性预测值为97.9%,总一致率为96.2%。对两种方法进行一致性Kappa检验,K=0.729(P0.01)。 结论：GenoType MTBDRplus方法检测结核分枝杆菌对利福平的耐药性,与传统比例法检测比较其敏感度和特异度,具有较高的一致性。但其较传统比例法快速,为临床早期诊断耐药结核病提供快速检测依据,利于临床早期合理治疗。
[Abstract]:Objective: to detect rifampicin resistance of Mycobacterium tuberculosis by GenoType MTBDRplus and traditional proportional method. Genotype MTBDRplus was used to detect rpoB gene, which is closely related to rifampicin resistance. The test can be completed within 24 hours. The traditional proportional method takes about three months to complete the test, although the method is simple, economical and easy to popularize, and it is difficult to meet the clinical needs for the diagnosis and treatment of tuberculosis. Genotype MTBDRplus provides rapid detection basis for early diagnosis of drug-resistant tuberculosis and is beneficial to early and reasonable clinical treatment. Methods: from April 2011 to September 2011, sputum samples from smear positive patients in Tianjin Binhai New District Tuberculosis Prevention and Control Institute were collected. The resistance of Mycobacterium tuberculosis to rifampicin was detected by GenoType MTBDRplus and traditional proportional method, respectively. Genotype MTBDRplus was used to detect rpoB gene closely related to rifampicin resistance. It was speculated that the resistance of rifampicin to rifampicin was speculated. GenoType MTBDRplus method included the treatment of positive sputum samples by alkali treatment-neutralization centrifugal precipitation method, the extraction and amplification of DNA,PCR with GenoLyse kit, and the results were read out. The rifampicin resistance of Mycobacterium tuberculosis was detected by traditional proportional method. Through the preparation of experimental materials, the suspension and dilution of bacteria were prepared, inoculated and cultured, and the results were reported. Kappa test is used to analyze the consistency of the two methods. The study was carried out in the reference room of Tianjin Tuberculosis Control Center. Results: 1. Among 55 smear positive specimens, 1 case was identified as non-tuberculosis mycobacteria (NTM), 1 case was contaminated, 1 case was positive and GenoType MTBDRplus could not be read. 2. 52 cases were compared with the proportional drug sensitivity test. 2. The result of rifampicin resistance. According to the results of proportional drug sensitivity, the sensitivity, specificity, positive predictive value, negative predictive value and total consistency rate of rifampicin resistance were 75%, 97.9%, 75%, 97.9% and 96.2%, respectively. the sensitivity, specificity, positive predictive value and negative predictive value of rifampicin were 75%, 97.9%, 75% and 97.9%, respectively. The consistency of the two methods was tested by Kappa, K 鈮，

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