Genotyping assays for the most common mutations associated with abacavir hypersen

与阿巴卡韦 Hypersen 相关的最常见突变的基因分型测定

• 批准号: 7684947
• 负责人: BERTRAND LEMIEUX
• 金额: $ 14.36万
• 依托单位: BIOHELIX CORPORATION
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-02-15 至 2009-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7684947
• 关键词: Abdominal Pain; Acquired Immunodeficiency Syndrome; Adult; Adverse reactions; Alleles; American; Anti-Retroviral Agents; Bathing; Biological Assay; Buffers; Caucasians; Caucasoid Race; Cells; Certification; Cheek structure; Clinical; Complex; Consensus; Containment; Cutaneous; DNA; DNA Sequence; Developed Countries; Developing Countries; Devices; Diagnostic; Diarrhea; Drug Interactions; Enzymes; Exanthema; Fever; Freeze Drying; Genotype; Gold; Guidelines; HIV; HIV-1; Heating; Highly Active Antiretroviral Therapy; Hour; Hypersensitivity; Incubated; Laboratories; Lamivudine; Lateral; Life; Malaise; Methods; Multi-Institutional Clinical Trial; Mutation; Names; Nausea and Vomiting; Nucleic Acids; Oligonucleotide Probes; Patients; Performance; Pharmaceutical Preparations; Phase; Process; Reaction; Reagent; Relative (related person); Research Personnel; Resistance profile; Resolution; Reverse Transcriptase Inhibitors; Risk; Sampling; Secure; Single Nucleotide Polymorphism; Specimen; Swab; Symptoms; System; Technology; Time; Treatment Protocols; Trizivir; Tube; United States; United States Dept. of Health and Human Services; Water; Zidovudine; abacavir; base; cost; design; food restriction; gastrointestinal symptom; genetic risk factor; helicase; migration; nucleoside analog; pill; point of care; prevent; prospective; public health relevance

DESCRIPTION (provided by applicant): We propose to develop a near patient assays to score 2 SNPs associated with abacavir-severe cutaneous adverse reactions (SCAR); i.e., HLA B*5701 and the Hsp70-Hom M493T allele (a.k.a. rs2227956CT). Nucleic acid templates will be isolated from cells obtained from cheek swabs with the Trimgen's BuccalQuick" (Sparks, MD). Two genotyping assays will be developed using our proprietary helicase dependent amplification (HDA). HDA products will be detected with a lateral flow device encased in an amplicon containment vessel that prevents contamination of the laboratory. In Phase I we will Implement design control, and secure ISO 13485 certification for BioHelix. We will also develop typing assays for the aforementioned SNP loci using this technology platform. Finally we will collaborate with PGXL to validate the performance of these assays relative to a gold standard (DNA sequencing) using a set of 50 clinical specimens collected by PGXL researchers. In Phase II, we would perform a prospective, multi-site clinical study aimed at validating the assays as a means of predicting the risk of SJS and TEN in patients being prescribed abacavir for the treatment of AIDS. PUBLIC HEALTH RELEVANCE: Abacavir (ABC), a human immunodeficiency virus-1 (HIV-1) nucleoside- analogue reverse transcriptase inhibitor, is coformulated with lamivudine (3TC ) and zidovudine (ZDV) in Trizivir; a single one a day pill with no food restrictions, limited drug-drug interactions, and a favorable resistance profile. Trizivir in combination with other HIV drugs, is recommended by the US Department of Health and Human Services Consensus Panel Guidelines as a possible initial treatment in antiretroviral-naive patients. ABV is also used in a combo with lamivudine in Epzicom(R). In addition, ABC (sold as a single drug under the name Ziagen(R)) is part of the first-line regimens used in developing countries because of its low cost (www.who.int/hiv/pub/guidelines/adult/en/index.html). Two single nucleotide polymorphisms (SNP) are highly association with abacavir-severe cutaneous adverse reactions (SCAR). HLA B*5701 is a known genetic risk factor for ABC hypersensitivity in Caucasians (Martin et al. 2005). The Hsp70-Hom M493T allele (a.k.a. rs2227956CT) in combination with HLA-B*5701 is strongly associated with ABC hypersensitivity. Although assays exist for HLA typing, these are costly, and yield more information that the strict necessary for detecting the risk of ABC hypersensitivity. For example, targeted DNA sequencing is used for high resolution typing, and sequence-specific primer (SSP), or sequence specific oligonucleotide probe (SSOP) based assays are used for low resolution HLA typing. SSP and SOPP assays typically 20 to 100 different primer sets or probes are used for each locus. DNA sequencing remains a complex and costly method for DNA diagnostics. The method we propose to develop would be lower cost and easy to implement in near patient settings.

描述（由申请人提供）：我们建议开发一种近患者测定法，以评分与阿巴卡维尔 - 严重性皮肤不良反应相关的2个SNP（疤痕）；即HLA B*5701和HSP70-HOM M493T等位基因（又称RS2227956CT）。 Nucleic acid templates will be isolated from cells obtained from cheek swabs with the Trimgen's BuccalQuick" (Sparks, MD). Two genotyping assays will be developed using our proprietary helicase dependent amplification (HDA). HDA products will be detected with a lateral flow device encased in an amplicon containment vessel that prevents contamination of the laboratory. In Phase I we will Implement design control,安全13485年生物helix的认证还将使用此技术平台开发上述SNP基因座的打字方法，我们将与PGXL合作，以使用PGXL研究人员收集的50个临床临床标本，以验证这些测定法的性能。作为一种预测SJS和十名患者的风险，被处方用于治疗公共卫生的相关性。每天只有一个没有食物限制的药丸，有限的药物相互作用以及有利的抗药性。 Trizivir与其他HIV药物结合使用，由美国卫生与公共服务部共识小组指南推荐，作为抗逆转录病毒患者的初始治疗方法。 ABV还与Epzicom（R）中的Lamivudine组合中使用。此外，由于其低成本，ABC（以Ziagen名称（r）名称出售为Ziagen（r））是使用的一线方案的一部分（www.who.int/hiv/pub/pub/guidelines/adult/adult/en/en/index.html）。两种单核苷酸多态性（SNP）与阿巴卡维尔 - 严重的皮肤不良反应（疤痕）高度关联。 HLA B*5701是高加索人中ABC超敏反应的已知遗传危险因素（Martin等，2005）。与HLA-B*5701结合使用的HSP70-HOM M493T等位基因（又称RS2227956CT）与ABC超敏反应密切相关。尽管存在HLA打字的测定，但它们是昂贵的，并且会产生更多信息，这是检测ABC超敏反应风险所必需的严格信息。例如，靶向的DNA测序用于高分辨率键入，基于序列特异性引物（SSP）或基于序列的特定寡核苷酸探针（SSOP）测定法用于低分辨率HLA键入。 SSP和SOPP分析通常用于每个基因座的20至100个不同的引物集或探针。 DNA测序仍然是DNA诊断的复杂且昂贵的方法。我们建议开发的方法是较低的成本，并且在近乎患者的环境中易于实施。