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为提高标记效率基因定位的准确性，将维甲酸诱导食管癌细胞系ＥＣ８７１２通过消减杂交获得的一个能抑制食管癌细胞生长的ｃＤＮＡＲＡ５３８克隆，并将其定位于人染色体８ｑ区。方法采用改良的非同位素标记方法，使生物素标记程度提高，对染色体原位杂交信号检出率有明显的影响。结果采用两次标记的探针进行染色体原位杂交，可见明显的杂交信号，观察６０个中期分裂相，两条染色单体上均出现阳性信号的核型有１８个，检出率达３０％，而仅用单次标记的探针进行杂交时，信号检出率明显降低，观察６０个分裂相，出现杂交信号的分裂相仅有７个，且多位于染色体的一条单体上。结论使用双标记荧光原位杂交技术，增加了标记分子的掺入率，提高了检测的成功率，将分化相关基因ＲＡ５３８定位于人８号染色体的长臂２区。
In order to improve the accuracy of marker-efficient gene mapping, a cDNA RA538 clone, which inhibits the growth of esophageal cancer cells, was induced by retinoic acid-induced esophageal cancer cell line EC8712 through subtractive hybridization and localized to the human chromosome 8q region. Methods The modified non-isotopic labeling method was used to increase the biotin labeling level and had a significant effect on the detection rate of chromosome in situ hybridization signals. RESULTS: Two labeled probes were used for chromosome in situ hybridization. Obvious hybridization signals were observed. 60 metaphases were observed. There were 18 karyotypes with positive signals on both chromatids. The detection rate reached 30%. However, when the probes were hybridized with single-labeled probes, the signal detection rate was significantly reduced. Observing 60 dissociation phases, there were only 7 dissociation phases of hybridization signals, and they were mostly located on one monomer of the chromosome. Conclusion The use of dual-label fluorescence in situ hybridization (FISH) has increased the incorporation rate of labeling molecules and improved the success rate of detection. RA538, a gene involved in differentiation, was located in the long arm 2 region of human chromosome 8.