A Low-Cost Portable Multiplex Serum Molecular Test for HBV and HCV Detection

用于检测 HBV 和 HCV 的低成本便携式多重血清分子测试

基本信息

批准号：
8981624
负责人：
Wan Y. Shih
金额：
$ 30万
依托单位：
LENIMA FIELD DIAGNOSTICS, LLC
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-07-01 至 2017-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8981624
关键词：
Affect Antibodies Area Binding Blinded Blood Blood Screening Cessation of life Chronic Chronic Hepatitis B Chronic Hepatitis C Clinical Clostridium difficile Country DNA DNA amplification Detection Developing Countries Devices Diagnosis Diagnostic Enzymes Feces Film Fluorescent Probes Frequencies Genes Genetic Genetic screening method Goals Health Personnel Hepatitis B Hepatitis B Surface Antigens Hepatitis B Virus Hepatitis C Hepatitis C virus Immunoassay Individual Infection Label Lead Link Liver Magnesium Molecular Monitor Nucleic Acids Organ Organ Donor Organ Transplantation Patients Phase Polymerase Chain Reaction RNA RNA amplification Relative (related person)Resources Reverse Transcription Risk Sampling Serologic tests Serum Source Specificity Stress Surface Surface Antigens System Technology Testing Time Tissue Donors Toxin Training Transfusion Transplanted tissue Universities Viral Virus base co-infection cost hazard lead titanate novel point of care prototype public health relevance receptor sensor solid solution viral DNA viral RNA viral detection

项目摘要

DESCRIPTION (provided by applicant): Hepatitis B (HB) and hepatitis C (HC) are two of the most widespread infections worldwide caused by the blood-borne HB virus (HBV) and HC virus (HCV), respectively. About 240 million and 150 million people suffer from chronic HB (CHB) and chronic HC (CHC), respectively while about 780,000 and 350,000-500,000 people die each year due to the consequence of HB and HC, respectively. In highly endemic countries, co-infection of HBV and HCV is also common. HB and HC are also important hazards for healthcare workers and for patients needing transfusion or organ transplant as CHB and CHC are asymptomatic and patients and blood/tissue donors may have HB or HC without knowing it, posing risks to healthcare workers and patients needing transfusion or tissue transplant. Currently HB and HC are diagnosed with two separate sets of tests which can detect HB and HC only weeks or months after the initial infection. Quantitative PCR (qPCR) could detect both HB and HC within the first week of infection but they are expensive not suitable for use in low-resource countries where HB and HC are endemic. They are also two tests, adding up the cost. The goal of the proposed study is to develop a low-cost, rapid, easy-to-use, multiplex, and accurate serum test to detect both HBV DNA and HCV RNA in one test to diagnose HB and HC at point of care, in low-resource HB and HC endemic countries, and to screen for blood and organ supplies. (PbMg1/3Nb2/3O3)0.65(PbTiO3)0.35 (PMN-PT) piezoelectric plate sensor (PEPS) is a new sensor that uses relative resonance frequency shift, f/f to detect binding of a target to the receptor on the PEPS surface. Unique to PMN-PT PEPS is that the detection f/f is enhanced by 1000 times due to the unique polarization switching mechanism within the PMN- PT layer. As a result, PEPS has demonstrated sensitivity similar to PCR in DNA detection but without the need of DNA extraction and DNA amplification. Lenima Field Diagnostics will develop a 3-PEPS array with one PEPS detecting HBV DNA, a second PEPS detecting HCV RNA, a third PEPS serving as a negative control. Preliminary results indicate 100 viruses/ml analytical sensitivity fr PEPS HBV DNA detection of HBV spiked in serum as similar to the 115 viruses/ml sensitivity of qPCR. Using a similar strategy, PEPS has demonstrated 95% sensitivity and 95% specificity in a separate study of Clostridium difficile toxin B gene detection in 40 patients' stools. It is expected that we will successfully develop the multiplexed PEPS HBV and HCV test to achieve a milestone of >90% sensitivity and >90% specificity. In phase II, we will develop the clinical prototype that includes a portable detection system and disposable array PEPS for patient sample tests towards FDA approval.

描述（由申请人提供）：乙型肝炎 (HB) 和丙型肝炎 (HC) 是世界范围内最广泛的两种感染，由血源性 HB 病毒 (HBV) 和 HC 病毒 (HCV) 分别引起约 2.4 亿和 2.4 亿人。 1.5 亿人分别患有慢性 HB (CHB) 和慢性 HC (CHC)，而大约 780,000 人和每年有 350,000-500,000 人因 HB 和 HC 的后果而死亡，在高度流行的国家，HB 和 HCV 的混合感染对于医护人员和需要输血或治疗的患者来说也很常见。器官移植，因为 CHB 和 CHC 无症状，患者和血液/组织捐献者可能在不知情的情况下患有 HB 或 HC，这给需要的医护人员和患者带来风险目前，HB 和 HC 通过两组独立的检测进行诊断，仅在初次感染后数周或数月即可检测到 HB 和 HC，但定量 PCR (qPCR) 可以在感染的第一周内检测到 HB 和 HC。它们价格昂贵，不适合在 HB 和 HC 流行的资源匮乏国家使用。它们也是两种测试，增加了成本。拟议研究的目标是开发一种低成本、快速、易于使用的方法。使用，多重、准确的血清检测，可在一次检测中检测 HBV DNA 和 HCV RNA，以便在 HB 和 HC 资源匮乏的国家在护理点诊断 HB 和 HC，并筛查血液和器官供应 (PbMg1/3Nb2)。 /3O3)0.65(PbTiO3)0.35(PMN-PT)压电板传感器(PEPS)是一种利用相对共振频移的新型传感器， f/f 检测目标与 PEPS 表面受体的结合 PMN-PT PEPS 的独特之处在于，由于 PMN-PT 层内独特的偏振切换机制，检测 f/f 增强了 1000 倍。因此，PEPS 在 DNA 检测中表现出类似于 PCR 的灵敏度，但不需要 DNA 提取和 DNA 扩增，Lenima Field Diagnostics 将开发一种带有一个 PEPS 检测的 3-PEPS 阵列。 HBV DNA，第二个 PEPS 检测 HCV RNA，第三个 PEPS 作为阴性对照初步结果表明，PEPS HBV DNA 检测血清中掺入的 HBV 的分析灵敏度为 100 个病毒/ml，与 qPCR 的 115 个病毒/ml 灵敏度相似。使用类似的策略，PEPS 在 40 名患者粪便中艰难梭菌毒素 B 基因检测的单独研究中表现出 95% 的敏感性和 95% 的特异性。预计我们将成功开发多重 PEPS HBV 和 HCV 检测，以实现 >90% 敏感性和 >90% 特异性的里程碑。在第二阶段，我们将开发包括便携式检测系统和一次性阵列 PEPS 的临床原型。患者样本测试以获得 FDA 批准。