滚环扩增技术直接检测痰标本中结核分枝杆菌利福平敏感性的初步研究

发布时间：2018-12-26 16:23

【摘要】：目的评价滚环扩增技术(rolling circle amplification,RCA)对痰标本中结核分枝杆菌利福平敏感性的直接快速检测效果。方法选取首都医科大学附属北京胸科医院住院的初治和复治涂阳肺结核患者24h痰标本32份,采用RCA法、直接测序法及传统药敏试验绝对浓度法测定痰标本中结核分枝杆菌对利福平的敏感性,并进行了比较。结果 32份痰标本中结核分枝杆菌,采用RCA法24株可检测到rpo B基因突变,其中23株单位点突变(516位2株,526位12株,531位9株),1株516位和526位联合突变,与直接测序结果均一致。32份痰标本的RCA方法检测与DNA测序结果和药物敏感性试验进行对比,28株结果均一致。4株结果不一致,其中2株RCA及测序无突变,药敏结果显示为耐药;2株RCA及测序显示有突变,药敏结果显示为敏感。结论RCA法用于痰标本中直接检测结核分枝杆菌对利福平的敏感性,同直接测序法结果一致,同传统药敏试验相比,RCA法能够大幅度节省时间,具有很好的临床应用和推广前景。
[Abstract]:Objective to evaluate the direct and rapid detection of rifampicin sensitivity of Mycobacterium tuberculosis in sputum by rolling loop amplification technique (rolling circle amplification,RCA). Methods Thirty-two sputum samples of first-time and re-treated smear positive pulmonary tuberculosis patients were collected from Beijing chest Hospital affiliated to Capital Medical University for 24 hours. RCA method was used. The sensitivity of Mycobacterium tuberculosis to rifampicin in sputum samples was determined by direct sequencing and traditional drug sensitivity test. Results among 32 sputum specimens, 24 strains of rpo B gene were detected by RCA method. Among them, 23 strains were unit point mutations (516, 526, 12, 531, 9), and 1 strain was 516 and 526, respectively. 32 sputum samples were detected by RCA method and compared with DNA sequencing results and drug sensitivity test. The results of 28 strains of sputum samples were consistent with those of 4 strains, and 2 strains of RCA and sequencing had no mutation. The results of drug sensitivity showed drug resistance. The RCA and sequencing of the two strains showed mutation, and the drug sensitivity of the two strains was sensitive. Conclusion the results of direct detection of mycobacterium tuberculosis sensitivity to rifampicin in sputum by RCA method are consistent with the results of direct sequencing. Compared with traditional drug sensitivity test, RCA method can greatly save time and has a good prospect of clinical application and popularization.
【作者单位】：北京市结核病胸部肿瘤研究所;
【分类号】：R521;R440

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