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目的用 3种先进的技术 -TBAccuProbeDNA杂交法、INNO LIPATMRIF TB(LiPA)探针试验和rpoβDNA基因序列测定的前瞻性研究 ,以鉴定结核分枝杆菌复合物 (MTBC)和非结核分枝杆菌(NTM)及其成本效益。方法在BactecMGIT 96 0培养系统中将获得的 10 5例阳性培养物筛选出 70例MTBC和 35例NTM ,用 3种技术鉴定并检测其rpoβ基因序列突变位点。 结果TBAccuProbe杂交阳性率是 95 7% (6 7/70 ) ,LiPADNA探针阳性率是 98 6 % (6 9/70 ) ,rpoβDNA序列测定阳性率是 97 1% (6 8/70 )。 3种方法两两比较差异在统计学上均无显著意义 (P >0 0 5 )。每份阳性培养物花费时间 :TBAc cuProbe杂交法和LiPADNA探针法在 2 4小时内完成 ,全部完成需 6～ 37天 ,平均为 15天 ;rpoβ基因序列测定需 4～ 5天完成 ,全部完成需 8～ 4 0天 ,平均 17天。比传统的 4～ 8周鉴定时间减少了很多。每份标本所需费用 3种方法各为 12 0、194和 96元。在 35例NTM中 ,3种方法均为MTBC阴性 ,同时rpoβDNA序列测定鉴定了NTM的种类。结论TBAccuProbe杂交和LiPADNA探针操作简便 ,所需时间短 ,只能鉴定MTBC和NTM。但LiPADNA探针还能鉴定rpoβ基因位点突变 ;rpoβ基因序列测定所需时间稍长 ,要有特定仪器和专门人员 ,但所需费用较低 ,除能鉴定MTBC外 ,还能
OBJECTIVE: To identify MTBD and NTM by using three advanced technologies - TBAccuProbeDNA hybridization, INNO LIPATMRIF TB (LiPA) probe and rpoβDNA sequencing. ) And its cost-effectiveness. Methods Totally 70 MTBC and 35 NTM were screened from 105 positive samples in Bactec MG 9600 culture system. The mutation sites of rpoβ gene were identified and detected by three techniques. Results The positive rate of TBAccuProbe hybridization was 95 7% (67/70). The positive rate of LiPADNA probe was 98 6% (6 9/70). The positive rate of rpoβ DNA sequencing was 97 1% (6 8/70). There was no statistically significant difference between the three methods (P> 0.05). Each positive culture takes time: TBAc cuProbe hybridization and LiPADNA probe method within 24 hours to complete, all completed 6 to 37 days, an average of 15 days; rpoβ gene sequencing takes 4 to 5 days to complete, all completed Need 8 ~ 40 days, an average of 17 days. Than the traditional identification of 4 to 8 weeks to reduce the time a lot. The cost of each specimen 3 ways each of 12 0,194 and 96 yuan. In 35 NTMs, all 3 methods were negative for MTBC, and the rpoβ DNA sequencing identified the species of NTM. Conclusion TBAccuProbe hybridization and LiPADNA probe are easy to operate and require short time, and can only identify MTBC and NTM. However, the LiPADNA probe can also identify mutations in the rpoβ gene locus. The time required to sequence the rpoβ gene is slightly longer, requiring special instrumentation and specialized personnel, but at a lower cost. In addition to identifying the MTBC,