CORE--APPLIED MOLECULAR BIOLOGY HEMATOLOGY

核心--应用分子生物学 血液学

• 批准号: 6584647
• 负责人: SUJIT SHETH
• 金额: $ 19.88万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-04-01 至 2003-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6584647
• 关键词: biomedical facility; blood disorder diagnosis; blood tests; cell sorting; clinical research; disease carrier state; genotype; hematology; hemoglobin Ss; human genetic material tag; human subject; molecular pathology; polymerase chain reaction; prenatal diagnosis; rapid diagnosis; sickle cell anemia

The molecular biology and hematological laboratory support to all units of the Center. Besides standard hematological and biochemical testings, it offers, by DNA analysis, precise genetic classification of all patients. The number of S genes and alpha-globin genes are enumerated and the haplotypes of the beta-globin like gene cluster are determined to define with precision the genotype of the patients. A battery of hematological parameters, including the levels of adhesion molecules reflecting endothelial damage, will be measured. These will be correlated with clinical manifestations by the statistical core unit to derive prognostic factors. This unit provides prenatal diagnosis of sickle cell diseases to all who request it. This is the largest program of prenatal diagnosis of sickle cell disease in the world. This unit will sharpen diagnostic techniques for prenatal diagnosis, such as the use of allele-specific PCR for fast diagnosis and the analysis of the Variable Number of Tandem Repeats (VNTR) to ascertain the contamination by maternal cells of chorionic villi samples. A major undertaking of this unit will be the development of techniques to obtain the prenatal diagnosis from a sample of maternal blood. This study, in cooperation with Dr. D.Bianchi at the New England Medical Center, will utilize analysis of fetal normoblasts circulating in the maternal blood. Nucleated red cells will be first concentrated by our technique of ultri- centrifugation on discontinuous gradients of arabino-galactane. Fetal cells will then be isolated by fluorescent activated sorting, using fluorescein-conjugated antibodies to cytoplasmic gamma-globin. The diagnosis will be made by PCR and the origin of the cells verified by analysis of Y-linked genes and/or by the VNTR analysis.

分子生物学和血液学实验室支持所有单位 中心。 除了标准的血液学和生物化学测试，它还提供了通过DNA 分析所有病人的基因类型S的数量 基因和α-珠蛋白基因被列举， β-珠蛋白样基因簇被确定为精确地定义了 患者的基因型。一系列血液学参数，包括 反映内皮损伤的粘附分子水平， 测定了这些将与临床表现相关联， 统计核心单元，以得出预后因素。 该单位向所有世卫组织提供镰状细胞病的产前诊断。 要求它。这是最大的镰状产前诊断计划 世界上最严重的细胞疾病 该单位将提高产前诊断的诊断技术， 由于使用等位基因特异性PCR进行快速诊断和分析， 可变串联重复数（VNTR），以确定 绒毛样本被母体细胞污染。 该股的一项主要任务是开发技术， 从母体血液样本中获得产前诊断。这项研究， 与新英格兰医疗中心的D.比安奇博士合作， 利用母体血液中循环的胎儿正常成红细胞的分析。 有核红细胞将首先通过我们的超浓缩技术浓缩， 在阿拉伯半乳聚糖的不连续梯度上离心。胎儿 然后通过荧光激活分选分离细胞， 荧光素偶联的抗细胞质γ-珠蛋白的抗体。的 诊断将通过PCR进行，细胞的来源将通过 Y连锁基因分析和/或VNTR分析。