Novel Assay Development for CFTR Genotyping

描述（由申请人提供）：该项目的目标是在第一阶段和第二阶段开发和验证一种新的CFTR基因分型诊断平台，该平台提供封闭管，手动，廉价和快速的检测。美国医学遗传学学院（ACMG）和美国妇产科医师学院（ACOG）最近建议计划生孩子的夫妇进行囊性纤维化（CF）基因（CFTR）内的突变检测。因此，CFTR诊断测定存在真实的且不断增长的市场。目前用于CFTR基因分型的测试系统执行起来很复杂，存在扩增子携带问题，并且需要花费数小时来执行。有了这笔资金，我们将使用我们的新平台GENE-CODE 2.0开发CFTR基因型的测试系统。GENE-CODE 2.0采用了扩展的遗传信息系统（AEGIS），允许在PCR过程中位点特异性酶促掺入报告分子。初步数据表明，GENE-CODE 2.0可以有效地用于CFTR deltaF 508、凝血酶原和遗传性血色病等靶标的基因分型。在第一阶段，我们将在ACMG/ACOG推荐的CFTR突变集上单独设计和演示平台。在第二阶段，我们将开发一个完整的系统，同时分析所有ACMG/ACOG推荐的CFTR突变。此外，通过内部检测功能，该系统应允许制造商以超快方式升级序列特异性，以包括将出现的包括其他种族人群的其他靶标。

DESCRIPTION (provided by applicant): The goal of this project, over Phase I and Phase II, is to develop and validate a new diagnostic platform for CFTR genotyping that provides closed tube, hands off, inexpensive and rapid testing. The American College of Medical Genetics (ACMG) and American College of Obstetricians and Gynecologists (ACOG) has recently recommended that couples planning to have a child be tested for mutations within the cystic fibrosis (CF) gene (CFTR). Thus, there is a real and growing market for a CFTR diagnostic assay. Today's testing systems for CFTR genotyping are complicated to perform, have problems with amplicon carry-over, and take hours to perform. With this grant funding, we will develop a testing system for CFTR genotypes using our novel platform GENE-CODE 2.0. GENE-CODE 2.0 employs an expanded genetic information system (AEGIS) that allows for site-specific enzymatic incorporation of reporter molecules during PCR. Preliminary data suggests that GENE-CODE 2.0 can be used effectively to genotype such targets as CFTR deltaF508, prothrombin and hereditary hemachromatosis. In Phase I we will design and demonstrate the platform on the ACMG/ACOG recommended set of CFTR mutations individually. In Phase II we will develop a complete system to analyze all ACMG/ACOG recommended CFTR mutations simultaneously. In addition, with internal assay capabilities, the system should allow the manufacturer to upgrade sequence specificity in an ultra-fast manner to include additional targets that will arise to include additional ethnic populations.