本文選題：納米基底片 + CD��； 參考：《廣東醫(yī)學(xué)》2017年02期

【摘要】：目的建立一種基于納米技術(shù)從孕婦外周血中分離胎兒有核紅細(xì)胞并進(jìn)行無(wú)創(chuàng)性產(chǎn)前診斷的方法。方法 Percoll 1.077單密度梯度離心法之后,選用抗體CD147修飾到以玻璃為載體的納米羥基磷灰石基底上,對(duì)新生兒臍血中的胎兒有核紅細(xì)胞進(jìn)行捕獲,利用胎兒特異性抗體HbF、紅細(xì)胞表面特異性抗體CD71以及細(xì)胞核染色劑DAPI對(duì)捕獲到的新生兒臍血中有核紅細(xì)胞進(jìn)行染色標(biāo)記,進(jìn)而實(shí)現(xiàn)分選。對(duì)正常不同孕周孕婦外周血、未孕健康婦女外周血及新生兒臍血中抗體CD147捕獲到的有核紅細(xì)胞進(jìn)行染色標(biāo)記分選。抽取10例行常規(guī)產(chǎn)檢時(shí)發(fā)現(xiàn)高危因素的孕婦外周血2 m L,用抗體CD147捕獲胎兒有核紅細(xì)胞并用X、Y、18、21染色體探針進(jìn)行熒光原位雜交。結(jié)果用HbF+CD71雙標(biāo)記染色法的胎兒有核紅細(xì)胞陽(yáng)性率與HbF單染和CD71單染標(biāo)記的細(xì)胞陽(yáng)性率相比差異有統(tǒng)計(jì)學(xué)意義(P0.05);HbF單染和CD71單染標(biāo)記的陽(yáng)性細(xì)胞率之間相比差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05);在孕16~20周時(shí)孕婦外周血中陽(yáng)性細(xì)胞數(shù)目達(dá)到頂峰,之后隨孕周增加而呈下降趨勢(shì)。10例高危孕婦外周血標(biāo)本均成功分離出胎兒有核紅細(xì)胞,并經(jīng)過(guò)FISH鑒定,本方法對(duì)男性胎兒診斷的靈敏度為85.71%,特異度為100.00%,對(duì)女性胎兒診斷的靈敏度為100.00%,特異度為85.71%,同時(shí)診斷出1例非整倍染色體胎兒,所有結(jié)果均經(jīng)后期隨訪證實(shí)。結(jié)論用納米材料為基底,抗體CD147捕獲,HbF+CD71雙染色標(biāo)記法對(duì)孕婦外周血中的胎兒有核紅細(xì)胞進(jìn)行識(shí)別和富集,可提高胎兒有核紅細(xì)胞的檢出率,可為進(jìn)一步產(chǎn)前診斷奠定基礎(chǔ),最佳時(shí)機(jī)應(yīng)該在孕16~20周。
[Abstract]:Objective to establish a non-invasive prenatal diagnosis method for the separation of fetal nucleated red blood cells from maternal peripheral blood based on nanotechnology. Methods after percoll 1.077 single density gradient centrifugation, antibody CD147 was modified onto the nano-hydroxyapatite substrate with glass carrier to capture fetal nucleated red blood cells from umbilical cord blood of newborn. Fetal specific antibody HbF, erythrocyte surface specific antibody CD71 and nuclear staining agent DAPI were used to stain and label nuclear red blood cells in captured umbilical cord blood. The nucleated red blood cells captured by CD147 in the peripheral blood of normal pregnant women with different gestational weeks and those of healthy women without pregnancy and umbilical cord blood of newborns were stained and sorted. Ten maternal peripheral blood samples with high risk factors were collected from 10 routine birth tests. Fetal nucleated red blood cells were captured by antibody CD147 and fluorescence in situ hybridization was performed with the XMY 1821 chromosome probe. Results the positive rate of fetal nucleated red blood cells using HbF CD71 double labeling method was significantly different from that of HBF single staining and CD71 labeled cells (P0.05) there was no difference between the positive rates of HBF single staining and CD71 single staining positive cells. The number of positive cells in peripheral blood of pregnant women reached the peak at 1620 weeks of gestation (P0.05). The fetal nucleated erythrocytes were isolated from peripheral blood samples of 10 high-risk pregnant women and identified by fish. The sensitivity and specificity of this method for the diagnosis of male fetus were 85.71 and 100.000.The sensitivity and specificity for female fetus were 100.00and 85.71respectively, and a case of aneuploid chromosome fetus was diagnosed. All the results were confirmed by late follow-up. Conclusion using nanomaterials as the substrate, the detection rate of fetal nucleated red blood cells in maternal peripheral blood can be improved by double staining method of antibody CD147 capture HbF CD71, which can lay a foundation for further prenatal diagnosis. The best time to get pregnant is 16 to 20 weeks.
【作者單位】： 承德市中心醫(yī)院產(chǎn)科;
【分類號(hào)】：R714.5

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