探针熔解曲线法快速检测结核分枝杆菌注射类二线药耐药突变

发布时间：2018-03-17 02:28

本文选题：结核分枝杆菌　切入点：注射类二线药　出处：《中国人兽共患病学报》2015年05期 　论文类型：期刊论文

【摘要】：目的评价结核分枝杆菌注射类二线药耐药突变检测试剂盒MeltPro TB/SL的分析性能以及临床性能。方法首先使用企业参考品对试剂盒的突变检出能力进行考察,之后将野生型基因组DNA以10倍梯度稀释后考察试剂盒的最低检测限,使用野生型质粒以及含常见耐药突变的质粒以不同突变比例混合考察试剂盒检测不均一耐药样本的能力,并使用23株非结核分枝杆菌标准株考察试剂盒的分析特异性,最后使用241份临床分离株对试剂盒进行临床评价。结果该试剂盒可以将9种耐药相关突变与1种野生型多态性与野生型进行区分。可以检测到低至5个拷贝的野生型基因组DNA。当突变比例为30%或更高时,试剂盒可以将其与野生型进行区分。23种非结核分枝杆菌的结果显示该试剂盒有良好的特异性。和比例法药敏试验进行对比,该试剂盒的临床灵敏度为卡那霉素90.0%(9/10),阿米卡星80.0%(8/10),卷曲霉素85.7%(6/7);特异性为卡那霉素98.8%(85/86),阿米卡星98.8%(85/86),卷曲霉素96.6%(86/89)。试剂盒检测结果与测序结果一致。结论该试剂盒灵敏度高、特异性好,可以快速、准确地检测结核分枝杆菌注射类二线药常见的耐药突变,有望应用于临床。
[Abstract]:Objective to evaluate the analytical and clinical properties of MeltPro TB/SL kit for detection of second line drug resistance in Mycobacterium tuberculosis injection. After dilution of wild type genomic DNA with 10 times gradient, the minimum detection limit of the kit was examined. The ability of using wild type plasmids and plasmids containing common drug-resistant mutations to detect heterogeneous drug resistant samples was studied by mixing the kits with different mutation proportions. The specificity of the kit was investigated by using 23 standard strains of non-tuberculous Mycobacterium tuberculosis. Finally 241 clinical isolates were used to evaluate the kit. Results the kit could distinguish 9 drug-resistance related mutations from 1 wild type polymorphism and detect as low as 5 copies of the wild type. When the mutation ratio is 30% or higher, The results showed that the kit had good specificity and was compared with the proportional susceptibility test. The clinical sensitivity of the kit is as follows: kanamycin 90.010 / 10, amicacin 80.010 / 810, entramycin 85.7 / 875 / 7; specificity kanamycin 98.8885 / 86C, amicacin 98.88.885 / 86C, amicacin 98.88.85 / 860.The results of the kit are in agreement with the sequencing results. Conclusion this kit has high sensitivity and good specificity. It can be used for rapid and accurate detection of common drug resistance mutations in mycobacterium tuberculosis injection-like second-line drugs, which is expected to be used in clinical practice.
【作者单位】：厦门大学生命科学学院"细胞应激生物学"国家重点实验室分子诊断教育部工程研究中心;中国食品药品检定研究院结核病疫苗室;
【基金】：国家“十一五”重大专项(No.2008ZX10003-004)资助~~
【分类号】：R446.5;R52

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