CFLP BASED DETERMINATION OF HEPATITIS C VIRUS GENOTYPE

基于 CFLP 的丙型肝炎病毒基因型测定

• 批准号: 2734759
• 负责人: MARY A BROW
• 金额: $ 30.36万
• 依托单位: THIRD WAVE TECHNOLOGIES, INC.
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-07-01 至 1999-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2734759
• 关键词: biotechnology; gene mutation; genetic library; genotype; hepatitis C virus; human genetic material tag; linkage mapping; method development; molecular cloning; restriction fragment length polymorphism; virus genetics

Hepatitis C virus (HCV) is a widespread, persistent disease affecting 1- 2% of the world's population. Only a fraction of HCV affected individuals respond to interferon therapy. Viral genotype has emerged as a clinically significant predictor for determining response to interferon. This proposal describes the application of a novel DNA fingerprinting technology, Cleavase Fragment Length Polymorphism (CFLP) to identifying HCV genotype. The specific aims of Phase II are to generate a two-pronged CFLP HCV genotyping kit for rapid identification of a broad range of HCV types and subtypes. This kit will be suitable for analysis of the highly conserved 5'NCR subgenomic region as a follow-on to widely used HCV detection systems as well as for analysis of the NS-5b region for high-resolution genotyping. Signature elements of CFLP cleavage patterns will be used to identify genotype in uncharacterized samples. The Phase II proposal includes internal validation testing and beta site testing in two prominent reference labs. The CFLP HCV genotyping kit will be capable of discriminating HCV genotype with up to 98% certainty (for single base changes, >98% for the multiple polymorphisms that form the basis of genotyping) in less time and for a lower cost than existing methods. PROPOSED COMMERCIAL APPLICATION Genotypic identification of hepatitis C Virus (HCV) is a rapidly growing area of research and clinical practice. In addition to affecting responsiveness to interferon, HCV genotype is likely to be an important factor in the evaluation of future antiviral therapies. CFLP is a fast, accurate, and user-friendly method for HCV genotyping. By developing a two-pronged assay, we propose to create a product suitable for immediate clinical use in detecting commonly occurring genotypes (based on the 5'NCR subgenomic region) as well as a more far-reaching method for higher-resolution typing (based on the NS-5b region).

丙型肝炎病毒（HCV）是一种广泛存在的持续性疾病，影响1- 占世界人口的2%。 只有一小部分HCV感染 个体对干扰素治疗有反应。 病毒基因型已经出现 作为临床上有意义的预测因子， 干扰素 该提案描述了一种新型DNA的应用， 裂解酶片段长度多态性（CFLP） 来鉴定HCV基因型。 第二阶段的具体目标是 制备了一种快速鉴定的CFLP HCV基因分型试剂盒， 广泛的HCV类型和亚型。 这个工具包将适合 用于分析高度保守的5 'NCR亚基因组区域作为 继广泛使用的HCV检测系统之后， NS-5 b区域进行高分辨率基因分型。 标志性元素 CFLP切割模式将用于鉴定基因型， 未鉴定的样品。 第二阶段提案包括内部 验证测试和beta测试在两个站点中有突出的参考价值 labs. CFLP HCV基因分型试剂盒将能够区分HCV 基因型具有高达98%的确定性（对于单碱基变化，>98%对于 形成基因分型基础的多个多态性） 并且成本低于现有方法。 建议的商业应用 丙型肝炎病毒（HCV）基因型鉴定是一个快速增长的 研究和临床实践领域。 除了影响 HCV基因型可能是影响干扰素应答的重要因素。 评估未来抗病毒治疗的因素。 CFLP是一种快速， 准确，用户友好的HCV基因分型方法。 通过开发一个 双管齐下的分析，我们建议创建一个产品，适合立即 在检测常见基因型中的临床应用（基于 5 'NCR亚基因组区域）以及更深远的方法， 高分辨率分型（基于NS-5 b区域）。

项目成果

New Cleavase Fragment Length Polymorphism method improves the mutation detection assay.

新的切割酶片段长度多态性方法改进了突变检测分析。

• DOI: 10.2144/00282pf02
• 发表时间: 2000
• 期刊: BioTechniques
• 影响因子: 2.7
• 作者: Oldenburg,MC;Siebert,M
• 通讯作者: Siebert,M

Algorithmic approach to high-throughput molecular screening for alpha interferon-resistant genotypes in hepatitis C patients.

丙型肝炎患者α干扰素耐药基因型高通量分子筛查的算法方法。

• DOI: 10.1128/jcm.36.7.1895-1901.1998
• 发表时间: 1998
• 期刊: Journal of clinical microbiology
• 影响因子: 9.4
• 作者: Sreevatsan,S;Bookout,JB;Ringpis,FM;Pottathil,MR;Marshall,DJ;DeArruda,M;Murvine,C;Fors,L;Pottathil,RM;Barathur,RR
• 通讯作者: Barathur,RR