目的　建立一种简便、价廉、有效地在大片段中检测突变的限制酶切指纹 -单链构象多态性(restriction endonuclease fingerprinting- single strand conformation polymorphism,REF- SSCP)方法。方法　以耳聋患者基因组 DNA为模板 ,扩增 Cx2 6基因片段 ,通过对扩增产物进行单链构象多态性、限制酶切指纹 -单链构象多态性和 DNA测序 ,比较 REF- SSCP技术对大片段中突变的检出效率。结果　长度为 72 4bp的扩增产物 SSCP结果未显示差异 ,REF- SSCP显示 3种不同带型 ,经测序发现该基因中的 79G→ A突变 ,突变情况与 REF- SSCP带型完全吻合 ,检出率为 10 0 %。结论　所建立的 REF- SSCP技术适用于大样本量地检测大片段 DNA中的突变 OBJECTIVE To establish a restriction endonuclease-fingerprinting-single strand conformation polymorphism (REF-SSCP) method that is simple, inexpensive and effective for detecting mutations in large fragments. Methods The genomic DNA of deaf patients was used as a template to amplify the Cx2 6 gene fragment. Single-strand conformation polymorphism (PCR), restriction fingerprinting-single strand conformation polymorphism and DNA sequencing were compared. The results of REF-SSCP Detection of mutations in large fragments. Results There was no difference in the SSCP of amplified products with a length of 72.4 bp. REF-SSCP showed three different bands. The 79G → A mutation in the gene was found by sequencing. The mutation was completely consistent with the REF- The rate is 100%. Conclusion The established REF-SSCP technique is suitable for detecting large mutations in large fragments of DNA