石蜡包埋组织中麻风菌基因扩增试验初探

Gene Amplification Test of Mycobacterium Leprae in Paraffin－ Embedded Tissue

目的 建立用石蜡包埋组织中麻风菌 DNA进行基因扩增的试验。方法 用 Texpat试盒将石蜡切片脱蜡、破壁和释放麻风菌 DNA并用 100％乙醇沉淀纯化 DNA，用引物 RPOT(F)和 RPUT(R)进行 PCR。结果 细菌指数大于 1＋的 33份石蜡包埋标本， 28份 PCR阳性，细菌指数为 0的标本 PCR阴性。阴性对照呈阴性反应，阳性对照呈阳性反应，方法敏感度达 0.04 pg DNA水平。结论 用本法可从福尔马林固定的石蜡包埋组织切片中释放麻风菌 DNA，并获得满意基因扩增结果。

Objective To establish a polymerase chain reaction(PCR) method to amplify DNA of M.leprae from fixed and paraffin－ embedded tissue(FPET). Methods The DNA of M.leprae was released from FPET by using Texpat Kit and purified with 100％ alcohol. The primers RPOT(1) and RPUT(2) were used to conduct the PCR. Results A total of 32 samples were examined. Out of 32 samples with BI of more than 1＋ , 28 were positive for PCR. The PCR was negative in a sample with BI=0. The sensitivity of PCR reached a level of 0.04 pg DNA. Conclusion This PCR method is very useful for amplifying the DNA of M.leprae from FPET.