基因芯片技术在结核分支杆菌耐药突变基因检测中的应用

目的通过基因芯片检测系统，快速检测临床样品中结核分支杆菌耐药突变情况。方法根据结核分支杆菌标准株H37Rv序列，设计了覆盖rpoB、katG，inhA基因突变区的系列寡核苷酸探针，制作膜芯片，检测临床样品中结核分支杆菌基因突变情况，以此判断耐药结果。结果在305例临床病例中，共检出阳性病例125例，其中阳性敏感病例64例，阳性突变病例61例，阳性率为40．98％，在125例阳性样品中，共发现有8种突变类型，其中10例531L，占7．94％，19例315M，占阳性样品中总数的15．08％。结论PCR与膜芯片杂交技术可临床检测结核分支杆菌对利福平和异烟肼的耐药性，并具有快速、简便、敏感的特点。

Application of gene chip technology in detection of mutant genes of Mycobacterium tuberculosis drug resistance

Objective To develop a new membrane chip assay for rapid detection of rpoB, katG and inhA gene mutation in clinical rifamp-resistant and /or isoniazide-resistant Mycobacterium tuberculosis isolates.Methods According to the sequence of H37Rv strain, fifteen oligonucleotide probes were designed specifically to detect the mutant sequences in rpoB, katG and inhA gene to identify the result of the drug resistance.Results 125 cases (40.98%) of mutations were found in 305 randomizedly selected Mycobacterium tuberculosis strains, in which 64 were sensitive cases, and 61 were mutant ones. In 125 positive samples, 8 kinds of mutant types were found, in which 10 cases (7.94%) were 531L, 19 cases (15.08%) were 315M.Conclusion Membrane gene chip technology is a sensitive and specific method for rapid detection of mutations of rifampin-resistant and /or isoniazid-resistant M. tuberculosis, and may be applied for clinical detection of drug resistance.