Single Well MPCR Donor Screen to ID Blood Pathogens

单孔 MPCR 供体筛查以识别血液病原体

• 批准号: 6789191
• 负责人: RAYMOND RANKEN
• 金额: $ 9.4万
• 依托单位: IONIS PHARMACEUTICALS, INC.
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-04-01 至 2005-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6789191
• 关键词: bioengineering /biomedical engineering; biohazard detection; biomedical automation; biomedical equipment development; blood bank /supply contamination; communicable disease control; hepatitis B virus group; hepatitis C virus; high throughput technology; human T cell lymphotropic virus type 1; human T cell lymphotropic virus type 2; human immunodeficiency virus 1; human immunodeficiency virus 2; human tissue; mass spectrometry; patient safety /medical error; polymerase chain reaction; public health; robotics; technology /technique development

DESCRIPTION (provided by applicant): The screening of the blood supply by the blood bank is expensive and labor intensive with the current state of the art being Gen-Probe's multiplex assay that has the potential to detect only two viral types simultaneously. Multiplex PCR offers a cost-effective solution, which, with refinement and full automation would allow screening of the donated blood supply, The major hurdle in the use of multiplex PCR as a detection protocol is the ability to accurately assign the identity of any one product in a potential mixture of many products, We propose to develop a single technology platform medical device with the potential to diagnose a broad variety of viral transfusion-transmitted disease organisms which may infect the human blood supply. This proposal leverages technology based on a high throughput, automated robotic system which performs the steps of cell lysis, nucleic acid extraction and purification, RT-PCR and detection of the PCR products by mass spectrometry. The exact mass of each PCR product in a mixture is used to determine the base composition of each product and this composition uniquely identifies all the organisms present in the original sample. This system utilizes a single-well multiplex PCR assay that will detect a large number of pathogens simultaneously. In phase one we propose to design and test primers for six blood banking pathogens (HIV types1 and 2, HTLV types 1 and 2, HBV and HCV). We will optimize the detection of each virus type singly in clinically relevant specimens (blood, serum and plasma) using single and multiplexed primer pairs. We will finally optimize the simultaneous detection of multiple pathogens in a single tube assay using multiplexed primer pairs.

描述（由申请人提供）：血液库对血液供应进行筛查昂贵且劳动力密集，目前的最新状态是Gen-Probe的多重分析，有可能同时检测两种病毒类型。 Multiplex PCR提供了一种具有成本效益的解决方案，通过改进和完全自动化将允许筛选捐赠的血液供应，使用多路复用PCR作为检测方案的主要障碍是能够准确分配任何一种产品的身份在许多产品的潜在混合物中，我们建议开发单个技术平台的医疗设备，以诊断可能感染人类血液供应的各种病毒输血传播的疾病生物。该建议利用基于高通量的自动机器人系统的技术，该系统执行细胞裂解，核酸提取和纯化，RT-PCR以及通过质谱对PCR产物的检测。混合物中每种PCR产物的精确质量用于确定每种产品的碱基组成，并且该组合物独特地识别了原始样品中存在的所有生物。该系统利用单孔多重PCR分析，该测定法将同时检测大量病原体。在第一阶段，我们建议设计和测试六种血液库病原体（HIV型1和2，HTLV型1和2，HBV和HCV）。我们将使用单一和多路复用引物对在临床相关标本（血液，血清和血浆）中单独优化每种病毒类型的检测。我们最终将使用多路复用引物对在单个管测定中同时检测多个病原体的同时检测。