GJB2单杂合突变非综合征型耳聋患者GJB2序列长度检测
发布时间：2018-09-14 12:31
【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。
[Abstract]:Objective to detect the insertion or deletion of large fragments of GJB2 in patients with non-syndromic deafness by single heterozygosity mutation of GJB2, and to study the method of long chain PCR and electrophoresis of GJB2 sequence. Methods the length of GJB2 sequence was measured in 201 patients with non-syndromic deafness with GJB2 single heterozygosity mutation by using long chain PCR method, and 111 normal hearing adults in the control group. The length and quantity of PCR products were detected by two-step PCR, with 0.8% agarose gel electrophoresis by adjusting the amount of DNA template and the number of cycles. The width, sample addition, electrophoretic voltage, current and time of PCR were adjusted to obtain clear bands. If there is a large base insertion or deletion in the PCR product, the restriction endonuclease BamHI is used to perform the endonuclease reaction, and the approximate position of the insertion or deletion is preliminarily determined. Results in 201 patients with GJB2 single heterozygosity, the length of GJB2 sequence was not inserted or deleted. The length of GJB2 sequence in control group was not abnormal. Conclusion the detection of GJB2 sequence length in non-syndromic deafness patients with GJB2 single heterozygosity mutation is not abnormal.
【作者单位】： 解放军总医院耳鼻咽喉头颈外科 解放军耳鼻咽喉科研究所;武警总医院耳鼻咽喉头颈外科;解放军总医院海南分院耳鼻咽喉头颈外科;
【基金】：国家十二五支撑项目(2012BAI09B00,2012BAI12B01) 国家自然科学基金重点项目(81230020);国家自然科学基金面上项目(81371096,81371098);国家自然科学基金青年项目(30801285) 卫生部行业专项基金(201202005) 北京市自然科学基金面上项目(7132177,7122172) 北京市科技新星计划(2009B34,2010B081) 国家高技术研究发展计划(“863”,2012AA020101)
【分类号】：R764.43
[Abstract]:Objective to detect the insertion or deletion of large fragments of GJB2 in patients with non-syndromic deafness by single heterozygosity mutation of GJB2, and to study the method of long chain PCR and electrophoresis of GJB2 sequence. Methods the length of GJB2 sequence was measured in 201 patients with non-syndromic deafness with GJB2 single heterozygosity mutation by using long chain PCR method, and 111 normal hearing adults in the control group. The length and quantity of PCR products were detected by two-step PCR, with 0.8% agarose gel electrophoresis by adjusting the amount of DNA template and the number of cycles. The width, sample addition, electrophoretic voltage, current and time of PCR were adjusted to obtain clear bands. If there is a large base insertion or deletion in the PCR product, the restriction endonuclease BamHI is used to perform the endonuclease reaction, and the approximate position of the insertion or deletion is preliminarily determined. Results in 201 patients with GJB2 single heterozygosity, the length of GJB2 sequence was not inserted or deleted. The length of GJB2 sequence in control group was not abnormal. Conclusion the detection of GJB2 sequence length in non-syndromic deafness patients with GJB2 single heterozygosity mutation is not abnormal.
【作者单位】： 解放军总医院耳鼻咽喉头颈外科 解放军耳鼻咽喉科研究所;武警总医院耳鼻咽喉头颈外科;解放军总医院海南分院耳鼻咽喉头颈外科;
【基金】：国家十二五支撑项目(2012BAI09B00,2012BAI12B01) 国家自然科学基金重点项目(81230020);国家自然科学基金面上项目(81371096,81371098);国家自然科学基金青年项目(30801285) 卫生部行业专项基金(201202005) 北京市自然科学基金面上项目(7132177,7122172) 北京市科技新星计划(2009B34,2010B081) 国家高技术研究发展计划(“863”,2012AA020101)
【分类号】：R764.43
【参考文献】
相关期刊论文 前1条
1 于飞;韩东一;戴朴;康东洋;张昕;刘新;朱庆文;袁永一;孙R，
本文编号：2242717
本文链接：https://www.wllwen.com/yixuelunwen/yank/2242717.html
最近更新
教材专著
