CORE--APPLIED MOLECULAR BIOLOGY HEMATOLOGY

核心--应用分子生物学 血液学

• 批准号: 6466617
• 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: 2001
• 资助国家: 美国
• 起止时间: 2001-06-20 至 2002-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6466617
• 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.

为所有单位提供分子生物学和血液学实验室支持 中心。 除了标准的血液学和生化测试外，它还提供 DNA 测试 分析，对所有患者进行精确的基因分类。 S的数量 基因和α珠蛋白基因被枚举，并且单倍型 确定β-珠蛋白样基因簇以精确定义 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将是 测量。这些将与临床表现相关 得出预后因素的统计核心单元。 该单位为所有患有镰状细胞病的人提供产前诊断 请求它。这是目前最大的镰刀产前诊断项目 世界上的细胞疾病。 该单元将提高产前诊断的诊断技术，例如 使用等位基因特异性 PCR 进行快速诊断和分析 可变串联重复次数 (VNTR) 以确定 绒毛膜绒毛样本被母体细胞污染。 该单位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， 与新英格兰医学中心的 D.Bianchi 博士合作，将 利用母体血液中循环的胎儿正常成细胞的分析。 有核红细胞将首先通过我们的超滤技术浓缩 阿拉伯半乳烷的不连续梯度离心。胎 然后通过荧光激活分选来分离细胞，使用 针对细胞质伽马珠蛋白的荧光素偶联抗体。这 将通过 PCR 进行诊断，并通过以下方法验证细胞的来源 Y连锁基因分析和/或VNTR分析。