本文關(guān)鍵詞：遺傳性耳聾基因芯片檢測(cè)方法的建立　出處：《中國(guó)優(yōu)生與遺傳雜志》2016年11期 　論文類型：期刊論文

  更多相關(guān)文章： 耳聾 基因突變 膜芯片 基因診斷

【摘要】：目的以膜芯片技術(shù)為平臺(tái),建立一種遺傳性耳聾基因芯片檢測(cè)方法(PCR-反向點(diǎn)雜交法)。方法根據(jù)近期在我國(guó)國(guó)內(nèi)開展的大規(guī)模耳聾分子流行病學(xué)調(diào)查結(jié)果及Gene Bank上報(bào)道的基因信息,設(shè)計(jì)并合成20條探針及5對(duì)引物檢測(cè)與遺傳性耳聾相關(guān)的四個(gè)熱點(diǎn)基因(GJB2、GJB3、SLC26A4和12S r RNA)中的10個(gè)突變位點(diǎn),10條探針點(diǎn)至尼龍膜上制成反向雜交膜芯片,應(yīng)用五重PCR技術(shù)一管完成所有位點(diǎn)的擴(kuò)增,建立了一種一次實(shí)驗(yàn)就可以檢測(cè)非綜合征遺傳性耳聾四個(gè)熱點(diǎn)基因上10個(gè)位點(diǎn)突變的方法。結(jié)果遺傳性耳聾基因芯片檢測(cè)方法對(duì)10種基因型檢測(cè)的靈敏度為10ng/μL;除肝素抗凝劑外,EDTA和枸櫞酸鈉抗凝劑都不是本檢測(cè)方法的干擾物質(zhì),溶血、脂血和黃疸樣本都不影響本方法的檢測(cè)結(jié)果;應(yīng)用這種方法檢測(cè)臨床樣本,檢測(cè)結(jié)果與測(cè)序結(jié)果比較,陽(yáng)性符合率為100%,陰性符合率為100%,總符合率為100%,經(jīng)χ2檢驗(yàn)分析,遺傳性耳聾基因檢測(cè)方法與測(cè)序法沒有明顯差別;重復(fù)性測(cè)試結(jié)果顯示在不同的實(shí)驗(yàn)條件下檢測(cè)結(jié)果完全一致,該方法能穩(wěn)定重復(fù)檢出。結(jié)論成功的建立了一種靈敏度高、特異性好、檢測(cè)結(jié)果穩(wěn)定、可靠、通量較高的遺傳性耳聾基因芯片檢測(cè)方法。
[Abstract]:Objective to take the membrane chip technology as the platform. Establishment of a method for genetic Deafness Gene Chip Detection by PCR- reverse Dot Hybridization (PCR- reverse Dot Hybridization). Methods according to the results of molecular epidemiological investigation on deafness and the gene information reported on Gene Bank in China recently. 20 probes and 5 pairs of primers were designed and synthesized to detect four hot genes associated with hereditary deafness: GJB2 and GJB3. Ten mutation sites of SLC26A4 and 12s r RNAs were used to produce reverse hybridization membrane chip on nylon membrane. All the sites were amplified by using a single tube of quintuple PCR technique. A method was established to detect 10 locus mutations in four hot spots of non-syndromic hereditary deafness. Results the sensitivity of the microarray method for detecting 10 genotypes of hereditary deafness was as follows: 1. 10ng / 渭 L; Besides heparin anticoagulant EDTA and sodium citrate were not interfering substances in this method. Hemolysis lipid blood and jaundice samples did not affect the results of this method. Compared with the results of sequencing, the positive coincidence rate was 100, the negative coincidence rate was 100, and the total coincidence rate was 100. The results were analyzed by 蠂 2 test. There was no significant difference between the methods of gene detection and sequencing of hereditary deafness. The results of repeatability test showed that the detection results were completely consistent under different experimental conditions, and the method could be detected stably and repeatedly. Conclusion A new method with high sensitivity, good specificity, stable and reliable detection results has been established successfully. A high throughput genetic microarray method for the detection of hereditary deafness.
【作者單位】： 深圳市南山區(qū)婦幼保健院檢驗(yàn)科;
【基金】：深圳市南山區(qū)技術(shù)研發(fā)和創(chuàng)意設(shè)計(jì)項(xiàng)目(衛(wèi)生類)(南科研衛(wèi)2014020)
【分類號(hào)】：R764.43;R440
【正文快照】： 耳聾是人類最常見的致殘?jiān)蛑?嚴(yán)重影響著人類的生活質(zhì)量[1]。全球共有3.6億人存在不同程度的聽力障礙,我國(guó)現(xiàn)有聽力殘疾者2004萬(wàn)人,7歲以下的聾兒達(dá)80萬(wàn)人,每1000個(gè)新生兒中就有1~3名聾兒,我國(guó)每年新生聾兒達(dá)3萬(wàn)人,每年新發(fā)遲發(fā)性耳聾患兒超過8萬(wàn)人[2,3]。耳聾已經(jīng)成為我

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