摘要
目的探索聚合酶链反应(PCR)和地高辛系统检测 t(14;18)的特异性和敏感性。方法用该法检测了不同来源 DNA 标本中的 t(14;18)。结果 3株阳性对照细胞系、14例滤泡性非何杰金淋巴瘤(FNHL)中13例、4例弥漫性非何杰金淋巴瘤(DNHL)中2例的 DNA 内检出 bcl-2-JH 基因;阴性对照细胞系、正常人和其他肿瘤 DNA 中均未检出 t(14;18)。DNA 序列测定结果亦证实检出的阳性片段确实为 bcl-2-JH 基因。敏感性分析结果证实该法较单用 PCR 技术敏感1000～10000倍。结论 PCR 和地高辛系统确为敏感、特异、实用和对人体无害的检测 t(14;18)的方法。
Objective To determine if the polymerase chain reaction(PCR)and Dig system is spe- cific and sensitive.Methods We detect and analyse the t(14;18)in lysates of the formalin-fixed, paraffin-embeded or fresh freezing tissue's DNA samples from 20 lymphoma and 7 other tumour patients by PCR and Dig system.Results All DNA samples of the other tumour patients,normal and K562 cell line,were negative and 15 of 20(75%)lymphoma patients,including 13 of 14(93%)follicular and 2 of 4(50%)diffuse lymphoma,were positive for the translocation.The result of sequencing DNA also confirmed the amplified positive segement was real bcl-2-JH fusion gene.Moreover,the method is 1000 to 10000 fold more sensitive than either Southern blot restriction analysis or stainning the PCR products with eithidium bromide.Conclusion The PCR and Dig system for detecting t(14;18)is sen- sitive,specific and practical,as well as no harm to health.In addition,the research also suggested that detecting t(14;18)in rare malignant cells of biopsy or bone marrow by this method is very helpful for early diagnosis and monitoring clinic relapse in tumour patients with t(14;18).
