DNA固定技术对结核分枝杆菌PCR产物杂交结果影响的研究

DNA fixation technique on influence of hybridization results of PCR products of M.tuberculosis

目的 比较各种ＤＮＡ 固定方法对结核分枝杆菌ＰＣＲ 产物杂交结果的影响。方法 选取结核分枝杆菌ＰＣＲ 扩增产物并点于尼龙膜上，用紫外线、加热和碱液三种方法固定ＤＮＡ，然后以ＰＣＲ 产物作探针用增强化学发光标记试剂进行标记，并对三种不同方法固定的ＰＣＲ 产物杂交分析。结果 结核分枝杆菌ＤＮＡ固定方法对ＰＣＲ 扩增产物的检测结果具有较大的影响，碱液固定具有较好的敏感性和重复性。结论 各地结核分枝杆菌ＰＣＲ 实验室可考虑用碱液法对ＰＣＲ

Objective To compare influence of various DNA fixation methods on hybridization of PCR products of M.tuberculosis.Methods PCR products of M.tuberculosis were obtained,then diluted into different concentration and added to the nylon membrane.PCR products on the nylon membrane were fixed with UV crosslinking,oven baking and alkali fixation method,then hybridized and analysed.Results It was found that DNA fixation method had a great influence on detection of PCR products of M.tuberculosis.Alkali fixation had a good sensitivity and reproducibility.Conclusion All the results suggested that PCR laboratory of M.tuberculosis could consider using alkali fixation to PCR products,then hybridized and detected.