IN SITU HYBRIDIZATION IN PRENATAL DIAGNOSIS

该项目的目标是开发显著改进的自动化 用于检测最常见的细胞遗传学 通过子宫内膜穿刺或绒毛膜穿刺获得的间期细胞异常 绒毛取样（CVS）。在间期细胞上使用DNA探针避免了 常规细胞遗传学分析所需的培养。这可以减少 产前诊断的时间和费用，并允许延长测试， 一个更大的风险人群。这种自动化将是 当胎儿细胞可以从母体中分离时， 血液样本这将消除与以下方面相关的风险： 将胎儿非整倍体筛查扩展到几乎所有的 所有怀孕。 我们将荧光DNA探针标记和自动图像分析， 计数间期细胞中的13、18、21、X和Y染色体。我们将使用 (1)一个独特的新的自动化显微镜和（2）4芯片CCD相机， 最近开发的用于（3）校正数字化彩色图像的算法 对于光谱重叠，以及（4）创建点计数的无偏估计 比例，沿着（5）厚样品成像的新方法， 用于定位细胞边界。提到的五条染色体 对于绝大多数在子宫内膜穿刺术中观察到的活的非整倍体， 或CVS。快速非整倍体筛查和性别决定也将有所帮助 诊断新生儿有多个先天性异常或模棱两可 生殖器通过原位杂交进行间期细胞分析， 提供诊断是在24小时内，而48-72小时， 对外周血进行常规细胞遗传学检查， 穿刺术或CVS。 建议的商业应用：一旦技术开发出来， 并有资格进行常规应用，它们将被纳入 PSII的PowerGene细胞遗传学自动化设备产品线， 在新系统中，并作为已在使用的现有系统的升级， 细胞遗传学实验室

The goal of this project is to develop significantly improved automated techniques for detecting the most commonly occurring cytogenetic abnormalities in interphase cells obtained by amniocentesis or chorionic villus sampling (CVS). Using DNA probes on interphase cells avoids the culturing required from routine cytogenetic analysis. This can reduce the time and cost of prenatal diagnosis and permit extension of the test to a larger segment of the at-risk population. Such automation will be particularly important when fetal cells can be isolated from maternal blood samples. This would eliminate the risk associated with amniocentesis or CVS and extend fetal aneuploidy screening to virtually all pregnancies. We will fluorescent DNA probe labeling and automated image analysis to count chromosomes 13, 18, 21, X, and Y in interphase cells. We will use (1) a unique new automated microscope and (2) 4-chip CCD camera with recently developed algorithms for (3) correcting digitized color images for spectral overlap, and (4) creating unbiased estimates of dot count proportions, along with (5) new methods for thick specimen imaging and for locating the cell boundary. The five chromosomes mentioned account for the great majority of viable aneuploidies observed at amniocentesis or CVS. Rapid aneuploidy screening and sex determination will also assist diagnosis of newborns with multiple congenital anomalies or ambiguous genitalia. Interphase cell analysis by in situ hybridization could provide a diagnosis is under 24 hours, compared to 48-72 hours for conventional cytogenetics done on peripheral blood and 10-14 days for amniocentesis or CVS. PROPOSED COMMERCIAL APPLICATION: As soon as the techniques are developed and qualified for routine application, they will be incorporated into PSII's PowerGene product line of cytogenetics automation equipment, both in new systems and as an upgrade to existing systems already in use in cytogenetics labs.