异源双链分析法用于结核和牛分枝杆菌的鉴别  

Identification of M.bovis from M.tuberculosis by DHPLC and SURVEYOR Nuclease

作　　者：张霞[1] 赵德华[1] 秦殊[2] 石瑞如[2] 张国龙 

机构地区：[1]郑州大学第三附属医院,郑州450052 [2]河南省胸科医院 [3]河南省公共卫生医疗中心

出　　处：《医药论坛杂志》2013年第6期31-32,共2页Journal of Medical Forum

摘　　要：目的牛分枝杆菌存在pncA基因C169G和oxyR基因G285A突变,而结核分枝杆菌则无此突变,本研究利用DHPLC和SURVEYOR酶法测定这两个位点是否有突变,探讨异源双链分析法在结核分枝杆菌、牛分枝杆菌鉴别中的临床应用价值。方法 SURVEYOR酶法和DHPLC法分析结核分枝杆菌与牛分枝杆菌的pncA及oxyR基因。结果 DHPLC图谱和SURVEYOR酶法电泳图谱在牛分枝杆菌和结核分枝杆菌显著不同,很容易将二者区分开来。结论 DHPLC法和SURVEYOR酶法灵敏,简便,快速,在结核与牛分枝杆菌的鉴别中优于传统的培养鉴别方法。Objective It is very difficult to differentiate M.bovis from M.tuberculosis from the aspect of molecular microbiology because of more than 99% similarity of their genome sequence.This work is to differentiate these two mycobacteria by DHPLC and SURVEYOR Nuclease methods according to pncA gene C169G mutation and oxyR gene G285A difference of M.bovis,for exploring the application of these two new methods of gene mutation detection based on heteroduplex analysis.Methods PncA gene and oxyR gene of M.tb and M.bovis were analyzed by DHPLC and SURVEYOR Nuclease methods.Results It was very easy to judge whether a sample was M.tb or M.bovis from DHPLC profile or SURVEYOR electrophoresis bands.Conclusion DHPLC and SURVEYOR Nuclease methods are sensible,simple,rapid and may become new methods to help to differentiate M.bovis from M.tb.

关 键 词：高效液相色谱法 核酸内切酶 结核分枝杆菌 牛分枝杆菌 

分 类 号：R446.5[医药卫生—诊断学]