间接原位巢式PCR法在结核病与结节病鉴别中的应用  

Application of indirect in situ nested PCR in identification of tuberculosis and sarcoidosis

作　　者：赵兰[1] 李秋红[1] 郑卉[2] 张容轩[2] 李惠萍[1] 

机构地区：[1]同济大学附属上海肺科医院呼吸科,上海200433 [2]同济大学附属上海肺科医院病理科,上海200433

出　　处：《中华医院感染学杂志》2011年第18期3963-3965,共3页Chinese Journal of Nosocomiology

基　　金：上海市科学技术委员会资助项目(09411951500)

摘　　要：目的观察结核病和结节病病理组织中结核分枝杆菌DNA检出率及分布,探讨间接原位巢式聚合酶链反应(PCR)方法在结核病与结节病鉴别中的作用。方法以结核分枝杆菌内插入序列IS6110为扩增靶序列,用间接原位巢式PCR法,检测30例结核病和30例结节病石蜡包埋病理组织中结核分枝杆菌DNA,以抗酸染色阳性之结核病石蜡包埋病理组织作阳性对照,分别以鼠胎肺及抗酸染色阳性、原位杂交时无TB-DNA探针的石蜡包埋肺组织作阴性对照。结果光镜下观察,30例结核病组织标本中有28例阳性,阳性率为93.3%;30例结节病组织标本中有1例阳性,阳性率为3.3%,经统计学分析两组差异有统计学意义(P<0.01);从病理组织形态学方面观察,两组标本阳性结果中结核分枝杆菌DNA分布有所不同,结核病组中具有向郎罕氏巨细胞周围集中的特征,而结节病组中则无此特征。结论用间接原位巢式PCR法检测石蜡包埋病理组织中结核菌DNA,可作为鉴别结核病和结节病的较可靠且直观的方法之一。OBJECTIVE To observe the DNA detection rate and distribution of Mycobacterium tuberculosis in histopathologic tissues of patients with tuberculosis and sarcoidosis and to explore the application of indirect in situ nested PCR in identification of tuberculosis（TB） and sarcoidosis.METHODS Insertion sequence IS6110 of M.tuberculosis was amplified using indirect in situ nested PCR to detect M.tuberculosis DNA in paraffin-embedded histopathological tissues of 30 sarcoidosis patients and 30 TB patients,by comparing to the acid-fast staining positive control of paraffin-embedded pathological tuberculosis tissues and negative controls of rat fetal lungs and paraffin-embedded lung tissues that were positive in acid-fast staining but negative in situ nested PCR without using TB-DNA probe.RESULTS Tissues with brown-yellow granules under light microscopy were considered as positive.28（93.3%） out of the 30 tuberculosis samples were positive,while 1（3.3%） out of the 30 sarcoidosis samples were positive.The positive rates of the two types of samples differed significantly（P〈0.01）.In addition,their Mycobactium tuberculosis DNA distribution in the positive samples was different from the pathological morphological point of view：DNA was distributed around Langhans＇ giant cells in TB samples,but not in sarcoidosis samples.CONCLUSION Indirect in situ nested PCR is feasible for detection of tuberculosis DNA in paraffin-embedded pathological tissues and can be used as a relatively reliable and intuitive method to distinguish tuberculosis and sarcoidosis.

关 键 词：结核病 结节病 结核分枝杆菌 聚合酶链反应 

分 类 号：R375[医药卫生—病原生物学]