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As a preliminary step to preimplantation diagnosis of sickle cell disease in unfertilized eggs or 8-cell embryos of heterozygous parents, we established quality control for detection of the mutant and normal alleles of the beta-haemoglobin gene using single buccal cells. Efficient polymerase chain reaction (PCR) amplification of a 680 base pair sequence of the beta-globin gene spanning the site of the sickle cell mutation was obtained for 79 per cent of single heterozygous cells. In 71 per cent of cases, both alleles were detected. With this current efficiency, we predict that a clinical preimplantation diagnosis at the 8-cell embryo stage could be carried out safely and reliably for a couple at risk of transmitting sickle cell disease to their children. 相似文献
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Diana W. Bianchi MD Gretchen K. Zickwolf Melissa C. Yih Alan F. Flint Ossie H. Geifman Marlena S. Erikson John M. Williams 《黑龙江环境通报》1993,13(4):293-300
Fetal nucleated erythrocytes (NRBC) in maternal blood are a non-invasive source of fetal DNA for prenatal genetic screening. We compared the effectiveness of three monoclonal antibodies for the separation of fetal cells from maternal blood by flow sorting. Mononuclear blood cells from 49 healthy pregnant women were incubated with antibody to CD 71, CD 36, and/or glycophorin A (GPA), employed singly or in combination with each other. These monoclonal antibodies recognize surface antigens on haematopoietic precursor cells. Successful isolation of fetal cells was defined as detection of Y chromosomal sequences in maternal blood from women carrying male fetuses, with absence of Y sequences when female fetuses were carried. Thus, gender prediction accuracy was used as a measure of fetal cell separation. Using anti-CD 71 to isolate fetal cells, gender prediction was 57 per cent correct; with anti-CD 36, it was 88 per cent correct. Anti-GPA, an erythrocyte-specific antigen, used alone or in combination with anti-CD 71 or 36, improved gender prediction to 100 per cent. We conclude that antibody to GPA improves the retrieval of fetal NRBC from maternal blood, permitting genetic analysis by the polymerase chain reaction. 相似文献
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We report in detail two series of chorionic villus cultivation for prenatal chromosomal diagnosis. Chorionic villi were sampled from both first- and second-trimester pregnancies. One hundred cultures were treated with trypsin–EDTA for 2 h and collagenase overnight, (method A) and 100 were treated with trypsin–EDTA for 1 h and collagenase for 2 h (method B). Using short-term enzymatic digestion, the cultivation time was reduced from 14 days to 6 days. Sufficient amounts of metaphases of good quality were present in 93 per cent of primary cultures harvested in situ, whereas enough metaphases of sufficiently good quality were in most cases present only after subcultivation of the cultures using method A. The decrease in cultivation time obtained is probably due to a higher yield of viable cells in monocellular suspension, an increased attachment efficiency, and a more rapid attachment of single cells (within 24 h). 相似文献
15.
In vitro characteristics of human fetal cells have been investigated after chorionic villus sampling at the first trimester and amniocentesis at the second trimester of pregnancy. Light microscopy revealed heterogeneous morphology of cell types in both the chorionic villus culture and the amniotic fluid cultures. Based on the experiments performed, chorionic villus cells are more sensitive to pronase, trypsin, and versene during subculture and have a higher DNA content per single cell and release more [125I]-Beta-human chorionic gonadotropin into culture medium than those found in amniotic fluid cells. The practical applications of this study are discussed. 相似文献
16.
ASMs模型中异养菌减衰系数之间的关系及其测定 总被引:2,自引:0,他引:2
国际水协(IAWQ)推出的ASM1、ASM2、ASM2D号模型中描述微生物的衰亡过程采用了死亡-再生模式,而ASM3号模型中描述微生物的衰亡则采用了传统减衰模式,文章从两种减衰模式的机理出发,指出了两者之间的区别与联系,并详细推导了两种减衰模式中各参数之间的关系,以此得出了异养菌传统减衰系数bH'和死亡-再生减衰系数bH的测定计算方法。文中在20℃条件下,对以印染废水为主的实际污水处理厂活性污泥的异养菌减衰系数bH'和bH进行了测定,所得结果分别为0.20和0.50。 相似文献
17.
Yun-Ling Zheng Maryann Demaria Dongkai Zhen Theresa J. Vadnais Diana W. Bianchi MD 《黑龙江环境通报》1995,15(10):897-905
Monoclonal antibody to fetal haemoglobin (a2γy2) has been proposed as a fetal-specific reagent. We developed an intracellular staining protocol that combines fluorescein isothiocyanate or phycoerythrin conjugated anti-γ with the DNA binding dye Hoechst 33342 to identify and flow sort fetal erythroblasts from maternal blood. Our preliminary observations on anti-γ-positive cells sorted from four different pregnant women are described here, using fluorescence in situ hybridization (FISH) with chromosome-specific probes to identify fetal cells. Our data demonstrate that far fewer candidate fetal cells are sorted with this protocol than by current cell surface staining methods that employ the monoclonal antibody CD71. This results in increased fetal cell sorting purities. With this protocol, standard FISH techniques require modification due to the rigorous fixation with 4 per cent paraformaldehyde. Our initial data indicate the promise of this approach. 相似文献
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喷射浮选处理含油污水的研究 总被引:1,自引:0,他引:1
曲爱平 《石油化工环境保护》2000,(3):6-9
重点对喷射浮选处理含油污水进行了试验研究,结果表明浮选室内射流泵的没水深度对喷射选除油效果有较大的影响,并且只有在气泡量大并充满浮选室及表面相对平静的状态下,才儿得较好效果,为此,我们在浮选室内加入自制的增料,使兼并的气海重新得到分散,消耗了一部分动能,减缓了浮选室的扰动,使除在大提高,从而为研制新型高效的喷射浮选设备提供了一条新途径。 相似文献