【摘要】：目的通過對血清學(xué)試驗確認(rèn)Rh D陰性個體的RHD基因型檢測,研究不同類型遺傳學(xué)背景Rh D陰性個體與抗-D同種免疫的關(guān)系。重點探討各種Rh D變異型個體的獻(xiàn)血及輸血策略,為稀有血液的合理應(yīng)用提供理論依據(jù)。方法1、對2011年3月至2013年6月醫(yī)院送檢的Rh陰性個體,通過詢問輸血史和妊娠史排除沒有免疫史的個體。采用鹽水試管法和間接抗人球試驗(IAT)排除血清學(xué)試驗常規(guī)試驗陽性的個體。對孕婦的流產(chǎn)史及生育史進(jìn)行分類,研究流產(chǎn)史及生育史對抗-D產(chǎn)生的影響。2、采用微柱凝膠法篩查抗體,并通過譜細(xì)胞進(jìn)行抗體鑒定,確認(rèn)抗體特異性。檢測這些樣本的Rh表型,包括D、C、c、E和e抗原。3、采用序列特異性引物PCR(PCR-SSP)技術(shù)及DNA序列分析技術(shù),分析樣本的RHD基因。人類紅細(xì)胞RHD陰性鑒定基因檢測試劑盒可檢測的基因型包括Rh D陽性、Rh D陰性、Rh D-CE(2-9)-D、DVa(Hus)、DVI III型、弱D15型和DEL RHD1227A。結(jié)果1、本研究觀察有免疫史Rh D陰性個體共計336例。其中22例有輸血史的個體均為醫(yī)院在輸血前檢查時發(fā)現(xiàn)抗體篩選陽性的送檢標(biāo)本樣本,對其進(jìn)行抗體鑒定試驗確定抗體特異性均為抗-D。通過詢問病史患者都是以往輸用過Rh D陽性血液或不能明確所輸用血液具體種類的個體。314例有過既往妊娠史的Rh陰性孕婦中產(chǎn)生抗-D46例,其中G2P0孕婦96例,產(chǎn)生抗-D個體4例,陽性率4.2%;G2P1孕婦41例,產(chǎn)生抗-D個體6例,陽性率14.6%;G3P0孕婦125例,產(chǎn)生抗-D個體10例,陽性率8.0%;G3P1孕婦38例,產(chǎn)生抗-D個體14例,陽性率36.8%;妊娠超過三次的孕婦38例,產(chǎn)生抗-D個體14例,陽性率36.8%。2、通過Dia Clon Rh-Subgroups+k卡測得對336例IAT陰性標(biāo)本進(jìn)行Rh表型鑒定。Ccee的個體為185例(55.06%),Ccee個體119例(35.42%),CCee個體14例(4.17%),cc Ee個體11例(3.27%),Cc Ee個體7例(2.08%)。3、通過Dia Clon Rh-Subgroups+k卡測得68例產(chǎn)生抗-D的Rh D陰性個體中,Rh表型為ccee的個體為56例(82.3%),Ccee個體5例(7.4%),CCee個體4例(5.9%),cc Ee個體2例(2.9%),Cc Ee個體1例(1.5%)。其中表型為ccee的個體產(chǎn)生抗-D的比例最高,達(dá)82.3%。4、通過人類紅細(xì)胞RHD陰性鑒定基因檢測試劑盒對336份樣品進(jìn)行檢測,發(fā)現(xiàn)其中249例(74.1%)個體完全缺失RHD基因,l68例(20.2%)個體攜帶RHD1227A等位基因,19例(5.6%)攜帶RHD-CE(2-9)-D融合基因。5、68例RHD1227A個體Rh表型為ccee的個體為0例,Ccee個體56例(82.4%),CCee個體8例(14.2%),cc Ee個體1例(1.8%),Cc Ee個體3例(5.36%)。6、抗體鑒定確定產(chǎn)生Ig G抗-D的個體68例,其中63例(92.6%)完全缺失RHD基因,5例(7.4%)攜帶RHD-CE(2-9)-D融合基因,攜帶RHD1227A等位基因未檢出產(chǎn)生抗-D的個體。產(chǎn)生抗體的基因型均為完全缺失RHD基因和RHD-CE(2-9)-D融合基因個體,DEL RHD1227A個體沒有產(chǎn)生抗-D。結(jié)論血清學(xué)確認(rèn)Rh D陰性個體的RHD基因型具有多態(tài)性,北京地區(qū)主要以RHD基因缺失為主,其次為RHD-CE(2-9)-D型和DEL RHD1227A�？�-D的產(chǎn)生與Rh陰性個體受到D抗原刺激的數(shù)量有關(guān),妊娠和生育的次數(shù)與抗D的產(chǎn)生呈正相關(guān)。完全缺失RHD基因和RHD-CE(2-9)-D融合基因個體被D抗原免疫時有同種免疫風(fēng)險,作為受血者應(yīng)輸用陰性血。Del個體與C抗原有高度相關(guān)性,在中國漢族人群中最常見的DEL型RHD1227A個體沒有產(chǎn)生抗-D,或許可以輸用陽性血。
[Abstract]:Objective to identify the RHD genotype of Rh D negative individuals by serological test, and to study the relationship between Rh D negative individuals with different genetic backgrounds and anti -D homoimmunization. The strategy of blood donation and blood transfusion for various Rh D variant individuals was focused on. Method 1, from March 2011 to 2013, 6 The Rh negative individuals in the monthly hospital were excluded by the history of blood transfusion and the history of pregnancy. By using saline test tube and indirect anti human ball test (IAT), the individuals who were positive in the serological test were excluded. The abortion history and birth history of pregnant women were classified, and the effects of abortion history and birth history against -D were studied, and.2 was studied. The antibody was screened by the microcolumn gel method and the antibody specificity was identified through the spectral cells. The Rh phenotypes of these samples were detected, including D, C, C, E and e antigen.3. Sequence specific primers PCR (PCR-SSP) and DNA sequence analysis techniques were used to analyze the RHD gene of the samples. Human erythrocyte RHD negative identification gene detection kit could be detected. The genotypes included Rh D positive, Rh D negative, Rh D-CE (2-9) -D, DVa (Hus), DVI III type, weak D15 type and the result of 1. In this study, there were 336 cases of immunological history negative individuals. The test determined that the specificity of the antibody was anti -D. by inquiring the patient history of the patients who had previously been transfused with Rh D positive blood or the specific type of blood of the individuals who were unable to specify the specific type of blood in the past pregnancy history of Rh negative pregnant women. Among them, 96 cases of G2P0 pregnant women, 4 cases of anti -D individuals, the positive rate of 4.2%, 41 cases of G2P1 pregnant women, were produced. There were 6 cases of anti -D, positive rate of 14.6%, 125 cases of G3P0 pregnant women, 10 cases of anti -D individual, 8% positive rate, 38 cases of pregnant women, 14 cases of anti -D individuals, 36.8% of the positive rate, 38 cases of pregnant women over three times, 14 cases of anti -D individuals, positive rate 36.8%.2, and Rh phenotypic identification of IAT negative specimens of 336 cases were tested by Dia Clon Rh-Subgroups+k card. Rh phenotypes were identified for IAT negative specimens of 336 cases by Dia Clon Rh-Subgroups+k card There were 185 individuals (55.06%), 119 Ccee individuals (35.42%), 14 CCee individuals (4.17%), 11 CC Ee individuals (3.27%), 7 Cc Ee individuals (2.08%).3, and 68 cases of -D Rh (82.3%) by Dia Clon Rh-Subgroups+k card. 2.9%) 1 cases (1.5%) of Cc Ee individuals. Among them, the proportion of individuals with CCEE was highest, reaching 82.3%.4. 336 samples were detected by human erythrocyte RHD negative identification gene detection kit, and 249 cases (74.1%) were completely missing RHD gene, L68 case (20.2%) carrying RHD1227A allele, 19 (5.6%) carrying RHD-. The Rh phenotype of CE (2-9) -D fusion gene was 0, 56 of Ccee individuals (82.4%), 8 in CCee (14.2%), 1 in CC Ee individual (1.8%), 3 in Cc Ee (5.36%). RHD1227A alleles were not detected to produce anti -D individuals. All the genotypes produced by the antibody were completely missing RHD gene and RHD-CE (2-9) -D fusion gene, and DEL RHD1227A individuals did not produce a -D. conclusion that serology confirmed Rh D negative individuals. The production of HD-CE (2-9) -D and DEL RHD1227A. resistance to -D was related to the number of Rh negative individuals stimulated by D antigen. The number of pregnancy and fertility was positively related to the production of anti D. The total deletion of RHD and RHD-CE (2-9) -D fusion genes were immune to the same immune risk when immunized with D antigen. Highly correlated, in Chinese Han population, the most common type DEL RHD1227A individuals did not produce anti -D and may lose positive blood.
【學(xué)位授予單位】：中國人民解放軍軍事醫(yī)學(xué)科學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：R446.6

【相似文獻(xiàn)】

中國期刊全文數(shù)據(jù)庫 前10條

1 朱俊,劉達(dá)莊,陸震宇,向東,沈偉;紅細(xì)胞輸血同種免疫發(fā)生率的研究[J];臨床輸血與檢驗;2003年01期

2 Hackney D.N. ,Knudtson E.J. ,Rossi K.Q. ,R.W.O'Shaug hnessy,郭慧芳;妊娠并發(fā)抗C型同種免疫的治療[J];世界核心醫(yī)學(xué)期刊文摘(婦產(chǎn)科學(xué)分冊);2005年02期

3 張維煌;同種免疫性新生兒血小板減少性紫癜[J];國外醫(yī)學(xué).輸血及血液學(xué)分冊;1982年02期

4 Parer JT;常青;;嚴(yán)重的Rh同種免疫—目前子宮內(nèi)診斷和治療的方法[J];國外醫(yī)學(xué).婦產(chǎn)科學(xué)分冊;1989年04期

5 Andreu G;紫外線照射在防止HLA同種免疫中的作用[J];國外醫(yī)學(xué).輸血及血液學(xué)分冊;1993年03期

6 王曉萌;Rh同種免疫:早期嬰兒溶血增加[J];國外醫(yī)學(xué).輸血及血液學(xué)分冊;1998年03期

7 ;紅細(xì)胞輸血的同種免疫發(fā)生率的前瞻性研究[J];中國輸血雜志;2001年S1期

8 楊衛(wèi)紅;同種免疫在臨床輸血中的影響及對策[J];宜春學(xué)院學(xué)報;2003年06期

9 王同顯;馬保鳳;孫波;張魯平;萬旭;;輸血后人類白細(xì)胞抗原與紅細(xì)胞同種免疫關(guān)系初探[J];臨床血液學(xué)雜志(輸血與檢驗版);2009年01期

10 費世柏;抗-D、-C和-G的差異:同種免疫妊娠的臨床關(guān)聯(lián)[J];國外醫(yī)學(xué).輸血及血液學(xué)分冊;1998年05期

中國重要會議論文全文數(shù)據(jù)庫 前1條

1 成紅星;閆桂香;胡榮花;陳玫;;同種免疫致新生兒血小板減少性紫癜1例[A];中國輸血1999年年會暨紀(jì)念A(yù)BO血型發(fā)現(xiàn)100周年學(xué)術(shù)交流論文專輯[C];1999年

中國碩士學(xué)位論文全文數(shù)據(jù)庫 前1條

1 閆芳;RhD陰性個體遺傳多態(tài)性與抗-D同種免疫關(guān)系研究[D];中國人民解放軍軍事醫(yī)學(xué)科學(xué)院;2015年

，

本文編號：2145265