安全輸血(3)ppt課件

1、紅細(xì)胞血型免疫分子生物學(xué)研討和臨床運(yùn)用：平安輸血紅細(xì)胞血型ISBT規(guī)范分類、命名及表述組織血型和器官血型紅細(xì)胞ABO血型、基因、表型亞型RH血型：基因構(gòu)造、表型輸血前血型檢測輸血前血型血清學(xué)檢測程序(P125-163)血型檢測:ABO血型RH血型不規(guī)那么抗體檢測和鑒定:交叉配血:常規(guī)檢測正反定型不符及緣由ABO血型亞型獲得性B、T多凝集檢測原那么及結(jié)果處置常規(guī)方法試劑常規(guī)檢測抗體鑒定常規(guī)要求根本實(shí)驗(yàn)技術(shù)普通程序及簡單鑒定復(fù)雜抗體鑒定鹽水交叉配血實(shí)驗(yàn)不完全抗體配血實(shí)驗(yàn)抗體篩檢和交叉配血實(shí)驗(yàn)結(jié)果分析血型檢定(P126表15-1)正反定型不符及緣由實(shí)驗(yàn)者操作錯誤紅細(xì)胞標(biāo)本問題血清標(biāo)本血型檢定(P12

2、6表15-1)實(shí)驗(yàn)者操作錯誤假陽性： 離心速度時間過 試劑或細(xì)胞標(biāo)本鹽水污染 不干凈試管 .假陽性： 未參與抗體 不認(rèn)識溶血-為陽性反響 試劑失效或不合格試劑 抗體與細(xì)胞比例少 .血型檢定(P126表15-1)紅細(xì)胞標(biāo)本問題 相容但不同型輸血或骨髓移殖后， 弱抗原性:亞型、惡性腫瘤、老年人 獲得性和遺傳性多搜集T/Tn， 血清標(biāo)本ABOAg-中和抗體試劑， 本身凝集素包被本身紅細(xì)胞 .血型檢定(P126表15-1) 纖維蛋白凝塊 不規(guī)那么抗體 規(guī)那么抗體:免疫抑制病、免疫抑制 6月嬰兒、老年人 骨髓移植，大量輸血漿 .血清標(biāo)本治療病人、老人ABO血型血型血清學(xué)亞型及抗原變化 分類主要根據(jù) 血清

3、學(xué)特征：表15.2、P131 新的ABO亞型或變異型BA 多凝集紅細(xì)胞：表14-1、P119 獲得性B紅細(xì)胞ABO血型血型血清學(xué)亞型及抗原變化 分類主要根據(jù):1. 抗原性弱2. 正反定型-血清抗體反響強(qiáng)度3. 紅細(xì)胞H抗原性強(qiáng)度4. 唾液中ABH血型抗原量和中和才干5. 凝集情況：混和凝集或弱凝集6. 紅細(xì)胞吸收放散抗A、抗B才干臨床有意義抗體和無意義抗體完全抗體和不完全抗體血型檢測不完全抗體的檢測交叉配血輸血前血型血清學(xué)檢測要點(diǎn)1. 檢測程序：2. 抗體性質(zhì)：3. 檢測技術(shù)和方法：對完全抗體的檢測對不完全抗體的檢測或運(yùn)用完全抗體的檢測、運(yùn)用不完全抗體的檢測1、ABO血型由Ag-Ab兩者決議-

4、不能只檢“一半血型2、病理和生理情況-ABO血型正反檢測結(jié)果不一致3、單抗試劑質(zhì)量要求和平安輸血關(guān)系：多凝集-獲得性B4、血型定型只是輸血前血型血清學(xué)檢測程序一部分血型血清學(xué)檢測和平安輸血要點(diǎn)ABO血型1、D弱型-產(chǎn)生抗D2、D抗原陽性-獻(xiàn)血員和病人為D陽性或陰性？3、抗D試劑要求：效價？識別表位？4、血型血清學(xué)結(jié)果和分子生物學(xué)結(jié)果與平安輸血關(guān)系？血型血清學(xué)檢測和平安輸血要點(diǎn)RH血型血型血清學(xué)檢測和平安輸血要點(diǎn)ABO血型RH血型結(jié)論1、規(guī)范化、規(guī)范化檢測程序重要性！2、目前臨床問題：忽視對不完全抗體的檢測3、血型根底研討成果、新技術(shù)促進(jìn)平安輸血1、研討和認(rèn)識RhD抗原的構(gòu)造2、高特異性的血型試

5、劑3、用于細(xì)胞流式儀檢測胎母出血FMH4、用于ELISA對預(yù)防性抗D定量檢測5、用于新生兒溶血病的抗D預(yù)防性制劑 BRAD-3IgG3),BRAD-5(IgG1)6、engineered-“null“antibody“block functional active maternal antibodies. 基因工程制備的“無效抗體一對“功能活性的母體抗體的阻斷 (a McAb-Anti-D engineered by altering in the Fc region to render it incapable of reacting with effect cells.such a “nu

6、ll antibody molecule could protect against RhHDN by binding to fetal red cells and blocking the binding of functionally active maternal antibodies.) 運(yùn)用抗D單克隆抗體的意義RHD基因RCR定型Vox Sang 2000;78(suppl 2):083-089Site Directed Mutagenesis of the Human RhD Antigen: Molecular Basis of D Epitopes. Neil D.Avent.

7、Wendy Liu.Douglas Voak.D-negative phenotypes_two major mecharisms_at the genomic level.RHD gene deletionRHD(psendogene):37bp insertion and nonsense mutation in RHDgene.All weak D phenotypes-:caused by coding region regions in the RHDgene. allered D antigen expression (lack some D epitopes)(previous

8、dogma that weak D phenotype individuals have merely quartitative but not qualitative differerces in RhD antigon is incorrect.)Partial D phenotypes-1.majority of partial D phenotypes are “Knockonts where loss of epitopes. 2.few partial D phenotypes involve just one amino acid change. 3.RoHar phenotyp

9、e-small number of D epitopes. RHCE gene exon 5 is replaced with an RHD equivalent.Recent work has concluded that the terms weak and pmtial D phenotypes should be reploced by the termABERRANT-D,as all weak D phenotypes investigated illustrate a unique D phenotype.Wagner Blood 2000 95:2699-2708.Vox Sa

10、ng 2000;78 (suppl):079-082Monoclonal Antobodies to RhD-Development and used.Marion L Scott.Donglas Voak.The last work-shop clustered the 103 anti-D monoclonal antibodies studied into 24 specificity (epitope)gwaps.No one anti-D monoclonal antibody has been found that is capable of reacting with all p

11、artial D type. IN DLAGNOSTICSFor DORNOR typing:1.Type all presentations of RhD that could cause immunization in a D negative individual as D positive.2.Use afasingle monoclonal auti-D is therefore not acceptable.3.The most common stuategy is to use one high avidity IgM monoclonal anto-D that gives s

12、trong reactivity with normal D phenotype,D and Dbut does not react with Dalongside a polyclonal or other monoclonal reagent that has been specifically selected for detection of D and very weak D.For PATIENT typing:1.Weak D and partial D individual are typing as D negative,as they will then safely be

13、 given D negative blood.2.Detection of weak D by antigloblm tests with an IgG anti-D. (poly or monocloual)is not recommened for patient testing.because of the risk of false positives.3.Reagents used for routine patient typing should not detedt D _for the reason that D individuals lack much of the no

14、rmal D antigen,and therefore readily make potent anti-D when transfused with D positive blood.For REFERENCE laboratorips:panel of monoclonal anti-D with different epitopet specificities are invaluable-samples with aberrant D typing resutls or individual who type as RhD positive but have anti-D in th

15、eir seaum.Vox Sang 2000;78 (suppl):079-082McAb-anti-D in diagnostic labs to measure FMH by flow cytometry-more accurate than the Rleihauer Test-assay.ELLSA-biotinylated monoclonal IgG anti-D in a competitive A-European pharmacopoeia study-comparing this with traditional auto-analyzer method for anti

16、-D quantitation.Use of monoclonal anti-D for prophylaxis.1.depends not only on the specificity and avidity of the monoclonal antibody-via its Fv regions.2.but also its functional activity in interating with effector cells-via its Fc region.3.in UK two McAb-Anti-D-clinical trials.BRAD-3,BRAD-5.(IgG3 and IgG1 respectively)4.“nullantibody:engineered by attering targeted residues in the Fc region to render it in capatle of reacting with effector cells.“nullantibody pratect against RRHDSMP1RHCETelomere CentromereRHCECHybridRh boxSMP1RHCE10987654