环介导等温扩增技术可视化检测结核分枝杆菌

发布时间：2018-05-14 06:57

本文选题：结核分枝杆菌 + 环介导等温扩增　；参考：《生物医学工程学杂志》2017年01期

【摘要】：为了提升对结核分枝杆菌(MTB)检测的实用性,本文开展了采用环介导等温扩增方法(LAMP)对MTB进行可视化检测的研究。首先,根据MTB 16S r DNA序列设计LAMP引物。然后,收集临床痰液样本并进行相应处理。为了评价LAMP的特异度和灵敏度,本文用电泳产物进行检测并用钙黄绿素进行可视化验证。最后,以细菌培养结果为金标准,用SPSS 17.0软件比较培养结果与LAMP的一致性。结果显示,特异度实验中无非特异扩增现象,灵敏度实验中的检测极限为10拷贝。另外,可视化产物检测方法和电泳后的灵敏性相一致。进一步进行临床实用性评价,灵敏性为94.47%,特异性为90%,与细菌培养无统计学差异,结果一致性较好(P0.05)。综上,LAMP技术高效而可视,具有在设备匮乏的现场和基层医疗机构的应用前景。
[Abstract]:In order to improve the practicability of detection of Mycobacterium tuberculosis (MTB), a method of loop mediated isothermal amplification (Lamp) for visual detection of MTB was carried out in this paper. Firstly, LAMP primers were designed according to MTB 16s r DNA sequence. Then, clinical sputum samples were collected and treated accordingly. In order to evaluate the specificity and sensitivity of LAMP, electrophoretic products were used for detection and visual verification with calcium xanthophyllin. Finally, the results of bacterial culture were used as gold standard, and the consistency between the results of culture and LAMP was compared with that of LAMP by SPSS 17.0 software. The results showed that there was no non-specific amplification in the specificity test, and the detection limit in the sensitivity test was 10 copies. In addition, the visual product detection method is consistent with the sensitivity after electrophoresis. Further evaluation of clinical practicability showed that the sensitivity was 94.47 and the specificity was 90. There was no statistical difference between the two groups and the results were consistent with each other (P 0.05). The lamp technology is efficient and visible, and has the prospect of application in the field and primary medical institutions.
【作者单位】：承德医学院病原生物学实验中心;中国人民解放军白求恩国际和平医院全军肝病诊治中心;
【基金】：全军医药卫生科研基金项目(CBJ14C010)
【分类号】：R52;R440

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