目的 :制备检测 DMD基因常见易缺失外显子的核苷酸探针 ,通过反向斑点杂交试验验证其特异性 ,为初步研制 DMD基因诊断芯片作准备。方法 :从健康人外周静脉血白细胞提取基因组 DNA,应用经典 18对引物对 DMD基因常见易缺失外显子进行 PCR扩增。将扩增产物与 p GEM@- T Easy载体连接 ,转化 E.coli JM10 9感受态细胞。克隆目的片段 ,并作测序鉴定。以此为探针进行反向斑点杂交试验。结果 :序列分析表明 ,克隆片段代表 DMD基因 18个常见易缺失外显子 ;以这些片段为探针进行反向斑点杂交 ,其结果与 PCR相符。结论 :克隆的基因片段用作探针在反向斑点杂交试验中显示出较好的特异性 ,可用于 DMD基因常见缺失的检测 OBJECTIVE: To prepare nucleotide probes for detection of common deleterious exons of DMD gene and to verify their specificity by reverse dot blot hybridization in preparation for the preliminary diagnosis of DMD gene chip. Methods: Genomic DNA was extracted from healthy white blood cells of peripheral venous blood, and 18 pairs of primers were used to amplify the common deleterious exons of DMD gene. The amplified product was ligated into pMGT - T Easy vector and transformed into E. coli JM109 competent cells. The target fragment was cloned and sequenced. As a probe reverse blot hybridization test. Results: Sequence analysis showed that the cloned fragment represented 18 common deletions exons of DMD gene. The reverse dot blot was performed with these fragments as probe, and the result was consistent with PCR. Conclusion: The cloned gene fragment used as a probe shows good specificity in the reverse dot blot hybridization assay and can be used for the detection of common deletions of DMD gene