用间期核双标记荧光原位杂交技术检测BCL2原癌基因易位

目的 建立一个准确易行的诊断非何杰金氏淋巴瘤的实验室指标。方法 建立了以BCL2酵母人工染色体克隆yA153A6、以及免疫球蛋白重链基因（IgH)噬菌体克隆H24为探针的双标记荧光原位杂交技术，用以在间期核中检测BCL2原癌基因易位，结果 在滤泡性淋巴瘤细胞系SU-DHL-6细胞中，BCL2和IgH的杂交信号呈非随机分布，与一个BCL2等位基因易位到14号染色体IgH基因之上相对应，一个BCL2和一个IgH杂交信号始终紧密相连，而在正常淋巴细胞中，这两种基因的杂交信号至少保持1/10核直径的距离。结论 该方法客观明了，准确易行，不受标本中分裂相数量和质量的限制。可望成为临床诊断非何杰金氏淋巴瘤的常规手段。

Detection of BCL2 translocation in an interphase neucleus using fluorescence in situ hybridization strategy

OBJECTIVE: To establish a specific method to diagnose non-Hodgkin's lymphoma in clinic. METHODS: Fluorescence in situ hybridization (FISH) strategy capable of detecting t(14;18)(q32;q21) chromosome translocation in an interphase nucleus was developed using a YAC clone containing the BCL2 gene and a phage clone containing IgHC(mu) region as probes. In the SU-DHL-6 cells, the translocated BCL2 allele (red) is overlapped by one of the green signals from IgH phage probe, while in normal lymphocytes the least distance of the hybridization signals from two different probes was either equal or larger than one-tenth of the diameter of a nucleus. RESULTS: Based on the relative position of the signals from BCL2 and IgH probes the BCL2 translocation can be clearly detected in an interphase nucleus. The metaphase number and quality in samples, which could significantly restrict the reliability of cytogenetic analysis, can be ignored here. CONCLUSION: This method shows a potentiality in clinical diagnosis of non-Hodgkin's lymphoma because of its objectivity, reliability and simplicity.