摘要
目的建立一种快速、准确、简便的检测结核分枝杆菌的方法。方法异硫氰酸胍裂解、酚-氯仿抽提、异丙醇沉淀法直接抽提结核分枝杆菌rRNA,地高辛标记探针斑点杂交,酶呈色观察结果检测结核分枝杆菌,用该法检测76例临床标本。结果该法至少可检测1.6×102/m l个结核分枝杆菌;20种其它分枝杆菌和9种非分枝杆菌检测结果均为阴性;两份强阳性和弱阳性标本抽提结核分枝杆菌rRNA重复杂交5次,结果完全一致;46例肺结核病人痰标本检出阳性26例,30例非结核病人痰标本检测结果均为阴性。结论斑点杂交法检测结核分枝杆菌方法简便,无需特殊仪器,有较高的敏感度、特异性和良好的重复性,适合在基层医院推广使用。
Objective To establish a speedy,accurate and simplified method to detect mycobacterium tuberculosis. Methods To extract the rRNA of mycobacterium tuberculosis by guanidine thiocyanate splltting,phenol-chloroform extraction,isopropanol precipitating and proceeding dot-blotting with the probe emblematized by Digoxin,observing the results by enzyme presents color. Seventy-six clinical isolates were detected by this method. Result This method could detect mycobacterium tuberculosis of 1.6×10^2/ml at least. The results of twenty kinds of other mycobacteria and nine kinds of non-mycobacteria were all negative. The rRNA of two strong-positlve and wean-positive isolates were hybridized five times repeatedly ,the results were correspondent. Sputum of forty-six pulmonary tuberculosis were detected and the results of twentysix specimens were positive. The results of thirty non-pulmonary tuberculosis were all negative. Conclusion Using dotblotting to detect mycobacterium tuberculosis is simplified and don＇t need specific instrument,it has high sensitivity,specificity and good repetition. It is fitting for local hospital to use.
出处
《现代检验医学杂志》
CAS
2005年第6期32-33,共2页
Journal of Modern Laboratory Medicine
基金
2003年盐城市第二批科技发展计划项目(编号:YK200344-3)
