聚合酶链式反应检测石蜡切片中结核杆菌DNA的敏感性的评价

Evaluation of Sensitivity of PCR to Determination of Mycobacterium Tuberculosis DNA in Paraffin bedded Pathology Specimens

目的 本研究通过聚合酶链式反应检测结核病组织石蜡切片中的结核菌 ,并与抗酸染色及免疫组织化学染色作一比较 ,探讨此方法在结核病诊断中的敏感性。方法 1用聚合酶链反应检测结核菌特征性的长度为 32 0 b P的DNA序列 ;2应用多克隆抗体对结核菌进行免疫组化标记 ,用 SABC法检测组织中的结核杆菌 ;3传统的萋 -尼抗酸染色。结果 聚合酶链式反应检测石蜡切片中结核菌的阳性率 77.8% (2 1/ 2 7) ;免疫组化检测结核杆菌阳性率 83.3% (15 /18) ;抗酸染色检测结核菌阳性率 35 .4 % (40 / 113)。结论 在石蜡切片中诊断结核 ,如果抗酸染色和免疫组化这两种方法都无法确诊 ,配合 PCR检测 ,诊断率会大大提高。本实验也说明 PCR是一种安全、稳定、灵敏度高的好方法 ,为临床诊断结核提供了一个非常有效的诊断技术。

Objective To evaluate the sensitivity of polymerase Chain Reaction(PCR) to diagnosis of Mycobacterium tuberculosis(TB) in the formalin fixed paraffin bedded pathology specimens,and to compare it with antiacidic dye and immunohistochemistry methods.Methods ① The specific DNA fragment (320 bp) of Tuberculosis was amplified and detected by PCR technology;②Immunohistochemistry method through SABC was performed to mark the Tubercle bacilli;③Conventional ZiehlNeelsen method was carried out to dye the Tuberculosic bacilli.Results The positive rate in detection of the Tuberculosis was 77.8%(21/27) for PCR,8.3%(15/18) for immunohistochemistry method and 35% (40/113) for Ziehl Neelsen method.Conclusions Compared with the other methods,PCR technology was a more valuable and useful method for the detection of the Mycobacterium tuberculosis in the paraffin bedded pathology specimens, especially when combined with immunohistochemistry method when the bacilli variated too much and were less typical.