Application of high-throughput approaches in the study of complex disorders

静脉血栓形成（VT）是导致死亡和发病的主要原因之一，在美国每年约有300，000例住院治疗和50，000例死亡，非洲裔美国人的发病率为141/100，000（1.4/1000）、104/100000（1/1000）、55/100000（0.6/1000）和21/100000（0.2/1000）。我们的计算表明，同时使用一组11个基因检测增加了静脉血栓形成检测的阳性预测值至少30倍。我们已经为识别血栓形成倾向MERT的方法申请了专利，该方法目前正在评估中！0Proof of Principle！1用于在几个种族人群中预测和准确评估遗传性血栓形成倾向。该方法包括快速，同时筛选所有已知的145个静脉血栓形成相关的复发性突变和多态性的9个分子抗凝血酶III（AT III），蛋白C，蛋白S，纤维蛋白原，因子V（FV），凝血酶原（因子II），亚甲基四氢叶酸还原酶（MTHFR），血管紧张素1转换酶（ACE）和纤溶酶原激活物抑制剂-1（派-1）基因的阵列。我们设计了291个寡核苷酸25聚体探针点样到微阵列上，通过多重PCR方法在一次扩增反应中扩增出40个扩增子，覆盖了9个不同基因的变异序列。我们现在正在验证我们的方法的分析有效性。 设计用于筛选年龄相关性黄斑变性发展易感性的微阵列的类似方法（用于识别和测试年龄相关性黄斑变性的方法-MERT-ARMD）已获得专利。视网膜相关性黄斑变性（ARMD）是美国和发达国家65岁及以上人群中严重视力丧失的最常见原因。这是一种多因素的疾病。我们设计了一种MERT-ARMD，使用基于杂交的高密度寡核苷酸阵列技术，同时筛查16种分子（CFH、LOC 387715、BF、C2、ABCR、Fibulin 5、VMD 2、TLR 4、CX 3CR 1、CST 3、MnSOD、MEHE、对氧磷酶、APOE、APOVL 4和hemicentin-1基因）中105种已知的年龄相关性黄斑变性相关突变和多态性。

Venous thrombosis (VT) is one of the leading causes of mortality and morbidity resulting in approximately 300,000 hospitalizations and 50,000 fatalities per year in the United States with an incidence of 141 per 100 000 African-Americans (1.4 per 1000), 104 per 100 000 Caucasians (1 per 1000), 55/100 000 in Hispanics (0.6per 1000) and 21 per 100 000 Asian/Pacific Islanders (0.2 per 1000). Our calculations demonstrated that concurrent use of a panel of 11 genetic tests increases the positive predictive value of testing for venous thrombosis at least 30-fold. We have patented a Method Evolved for Recognition of Thrombophilia MERT that is presently under evaluation for !0proof of principle!1 for potential prediction and accurate assessment of hereditary thrombophilia in several ethnic populations. The methodology involves rapid, concurrent screening of an array of all known 145 venous thrombosis-associated recurrent mutations and polymorphisms in nine molecules antithrombin III (AT III), protein C, protein S, fibrinogen, factor V (FV), prothrombin (factor II), methylenetetrahydrofolate reductase (MTHFR), angiotensin 1-converting enzyme (ACE) and plasminogen activator inhibitor-1 (PAI-1) genes . We have designed 291 oligonucleotide 25mer probes to be spotted onto the microarray and achieved to amplify 40 amplicons covering the variation sequences from nine different genes in a single amplification reaction by Multiplex PCR assay. We are now in the process of verifying the analytic validity of our method. A similar approach for the design of a microarray screening for susceptibility to development of age-related macular degeneration (Method Evolved for Recognition and Testing of Age-Related Macular Degeneration-MERT-ARMD has been patented. Age-related macular degeneration (ARMD) is the most common cause of severe vision loss in the United States and developed countries among people 65 years of age and older. It is a multifactorial disorder. We have designed a MERT-ARMD that will concurrently screen 105 known age-related macular degeneration-associated mutations and polymorphisms in 16 molecules (CFH, LOC387715, BF, C2, ABCR, Fibulin 5, VMD2, TLR4, CX3CR1, CST3, MnSOD, MEHE, paraoxonase, APOE, ELOVL4 and hemicentin-1 genes), using hybridization-based, high-density oligonucleotide array technology.