【摘要】：目的檢測GJB2單雜合突變非綜合征型耳聾患者GJB2全序列中是否有大片段的堿基插入或缺失,研究GJB2全序列長鏈PCR方法和電泳方法。方法應用長鏈PCR方法對201例GJB2單雜合突變非綜合征型耳聾患者進行GJB2全序列長度檢測,對照組為111例聽力正常的成年人。應用兩步法PCR,調整加入DNA模板量、PCR延伸時間、循環(huán)次數(shù)等,0.8%瓊脂糖凝膠電泳檢測PCR產(chǎn)物的長度和量,調整加樣槽的寬度、加樣量、電泳電壓、電流、電泳時間得到清晰條帶,若PCR產(chǎn)物存在大片段堿基插入或缺失,用限制性內切酶BamHI進行內切酶反應,初步判斷插入或缺失的大致位置。結果 201例GJB2單雜合突變患者中,GJB2序列長度未見大片段堿基插入或缺失。對照組GJB2序列長度未見異常。結論 GJB2單雜合突變非綜合征型耳聾患者中GJB2序列長度檢測未見明顯異常。
[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.
【作者單位】： 解放軍總醫(yī)院耳鼻咽喉頭頸外科 解放軍耳鼻咽喉科研究所;武警總醫(yī)院耳鼻咽喉頭頸外科;解放軍總醫(yī)院海南分院耳鼻咽喉頭頸外科;
【基金】：國家十二五支撐項目(2012BAI09B00,2012BAI12B01) 國家自然科學基金重點項目(81230020);國家自然科學基金面上項目(81371096,81371098);國家自然科學基金青年項目(30801285) 衛(wèi)生部行業(yè)專項基金(201202005) 北京市自然科學基金面上項目(7132177,7122172) 北京市科技新星計劃(2009B34,2010B081) 國家高技術研究發(fā)展計劃(“863”,2012AA020101)
【分類號】：R764.43

【參考文獻】

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