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中文摘要：
目的检测GJB2单杂合突变非综合征型耳聋患者GJB2全序列中是否有大片段的碱基插入或缺失，研究GJB2全序列长链PCR方法和电泳方法。方法应用长链PCR方法对201例GJB2单杂合突变非综合征型耳聋患者进行GJB2全序列长度检测，对照组为111例听力正常的成年人。应用两步法PCR，调整加入DNA模板量、PCR延伸时间、循环次数等，0．8％琼脂糖凝胶电泳检测PCR产物的长度和量，调整加样槽的宽度、加样量、电泳电压、电流、电泳时间得到清晰条带，若PCR产物存在大片段碱基插入或缺失，用限制性内切酶BamHI进行内切酶反应，初步判断插入或缺失的大致位置。结果201例GJB2单杂合突变患者中，GJB2序列长度未见大片段碱基插入或缺失。对照组GJB2序列长度未见异常。结论GJB2单杂合突变非综合征型耳聋患者中GJB2序列长度检测未见明显异常。
英文摘要：
Objective To detect whether or not does inse~ion or deletion of large fragment nucleotides exist in GJB2 gene for the NSHI （nonsyndromic hearing impairment） patients with monoallelic GJB2 Gene Mutations, and to study long chain PCR method and electrophoresis for the full sequence length of GJB2 gene. Methods We detected the sequence length of GJB2 gene for 201 patients with monoallelic GJB2 gene mutations and 111 cases of control group with normal hearing by long chain PCR method. The product amount of PCR was adjusted by the amount of the DNA template, the extension time and the cycles of PCR. The sequence length of PCR product of the GJB2 gene was detected by 0.8% agarose gel electrophoresis. To ob- tain a clear electrophoresis strip, we changed the width of the well, the volume of sample of PCR product, the electrophoresis voltage and current, electrophoresis time, etc. If the sequence length of PCR products had obviously increase or decrease, there could exist insertion or deletion of large fragment nucleotide in the GJB2 gene sequence. We determined the approximate loca- tion of the insertion or deletion by restriction enzyme reaction of BamHI enzyme. Results We didn＇ t detect obvious increase or decrease of the sequence length of GJB2 gene in 201 patients with monoallelic GJB2 gene mutations and in the control group. Conclusion Insertion or deletion of large fragment nucleotide in GJB2 gene sequence was not detected in NSHI pa- tients with monoallelic GJB2 gene mutations.
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