Point of care detection of HPV in saliva

唾液中 HPV 的护理点检测

• 批准号: 10761543
• 负责人: ROYA KHOSRAVI-FAR
• 金额: $ 32.97万
• 依托单位: INNOTECH LLC
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-12 至 2024-09-11
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10761543
• 关键词: Base Sequence; Biochemical; Biological Assay; Biological Markers; Biosensor; Cancerous; Caring; Chemical Engineering; Clinical; Clustered Regularly Interspaced Short Palindromic Repeats; DNA; DNA amplification; Data; Dentists; Detection; Development; Devices; Diagnosis; Diagnostic; Disease; Early Diagnosis; Electrodes; Engineering; Ensure; Enzymes; Future; Genes; Genetic Materials; Goals; Gold; Guide RNA; HPV-High Risk; Housekeeping; Housekeeping Gene; Human Papillomavirus; Human papilloma virus infection; Human papillomavirus 16; Human papillomavirus 18; Incidence; Laboratories; Malignant Neoplasms; Massachusetts; Medical; Methods; Microfluidic Microchips; Microfluidics; Monitor; Monitoring for Recurrence; Morbidity - disease rate; Multiple Partners; Nucleic Acids; Oropharyngeal Squamous Cell Carcinoma; Pathologist; Patients; Performance; Personal Satisfaction; Phase; Plant Leaves; Point of Care Technology; Predictive Value; Preparation; Program Development; ROC Curve; Reagent; Recurrence; Resources; Reverse Transcriptase Polymerase Chain Reaction; Saliva; Sampling; Sensitivity and Specificity; Serum; Sexting; Signal Transduction; Small Business Innovation Research Grant; Sore Throat; Specimen; Symptoms; System; Technology; Test Result; Testing; Time; United States; United States National Institutes of Health; Validation; Viral; Virus Diseases; Work; amplification detection; assay development; commercialization; cost; design; detection assay; detection sensitivity; detection test; diagnostic assay; diagnostic platform; effective therapy; endonuclease; high risk; high risk population; improved; innovation; invention; malignant oropharynx neoplasm; manufacture; manufacturing process development; mortality; multiplex detection; non-invasive monitor; novel; nucleic acid delivery; nucleic acid detection; particle; plasmid DNA; point of care; point of care testing; point-of-care detection; precision medicine; process repeatability; product development; programs; promote resilience; prototype; relapse patients; screening; success; technology platform; translational medicine; treatment response; viral detection; virus genetics

Expanded point-of-care (POC) screening and monitoring options for detection of disease biomarkers from easily collectable specimens such as saliva are needed to reduce the incidence and mortality rates of debilitating diseases and enhance the resilience of the medical system, and improve care. InnoTech Precision Medicine with Massachusetts Institute of Technology is working to develop an innovative diagnostic platform that can be used for POC screening and monitoring of disease biomarkers. In our initial proof-of-concept, we are developing a POC test for detection of high-risk HPV in saliva. HPV is a viral infection that causes most Oropharyngeal squamous cell carcinoma (OPSCC) cases. Recent studies have demonstrated that HPV can be detected in saliva from patients with OPSCC. Currently, the initial screening and diagnosis of OPSCC is done at the late symptomatic stage based on symptoms that are often mild and can mimic less serious conditions such as cold or sore throat, delaying diagnosis and treatment. DNA tests for HPV are most often based on RT-PCR detection of the viral genetic material from serum or saliva that is collected and sent to central laboratories. RT-PCR-based tests rely on the amplification and detection of viral genetic material. These tests have long actionable turn- around-time, and high cost. Thus, there are no rapid, accurate and low-cost tests for routine monitoring of high- risk groups (i.e., young, sextually active, multiple partner) and monitoring of response to therapy and recurrence. Thus, a POC test for non-invasive monitoring of HPV in saliva during well-being exams can have a significant impact and save lives. Our objective is to develop a rapid, easy-to-use prototype for the detection of HPV in saliva based on the combination of biochemical signal amplification with CRISPR-Cas 12 electrochemical readout and our proprietary robust, accurate and low-cost gold-leaf electrodes. Utilizing our proprietary gold-leaf electrode along with a CRISPR-Cas12 assay for sensitive and specific electrochemical detection of target nucleic acid (NA) biomarkers, we have already demonstrated the detection of HPV 18 with plasmid DNA as well as in patient specimen. Our overall objective for this proposal is to develop a reproducible process for processing of saliva and develop a prototype electrochemical biosensor for the multiplex detection of HPV 16 and HPV 18 along with a reference gene to ensure accurate test results. To accomplish this goal, we will first develop a workflow for spatial multiplexing of our gold leaf electrodes for detection of HPV16, HPV 18 and an optimal reference gene. We will then verify this multiplexed test initially with plasmid DNA, and then with saliva specimens. We anticipate that combining easy-to-fabricate spatially multiplexed electrodes with CRISPR- Cas12a for the detection of a specific nucleic acid sequence will enable our Phase II development, regulatory approval and commercialization of a point-of-need technology that can be scaled and manufactured for accurate, easy-to-use and low-cost point-of-need testing and to reduce the burden of HPV infection and OPSCC.

扩大床旁（POC）筛查和监测选项，用于检测疾病生物标志物， 需要可收集的样本，如唾液，以减少衰弱性疾病的发病率和死亡率。 加强医疗系统的复原力，并改善护理。InnoTech精准医疗 与马萨诸塞州理工学院正在努力开发一个创新的诊断平台，可以 用于POC筛查和疾病生物标志物监测。在我们最初的概念验证中，我们正在开发 POC测试用于检测唾液中的高危HPV。HPV是一种病毒感染， 鳞状细胞癌（OPSCC）病例。最近的研究表明，HPV可以在 口腔鳞状细胞癌患者的唾液目前，OPSCC的初步筛查和诊断是在晚期进行的。 症状阶段，症状通常很轻微，可以模仿不太严重的情况，如感冒 或咽喉痛，延误诊断和治疗。HPV的DNA检测通常基于RT-PCR检测 收集并送往中心实验室的血清或唾液中的病毒遗传物质。基于RT-PCR 测试依赖于病毒遗传物质的扩增和检测。这些测试有很长的可操作性- 时间长，成本高。因此，没有快速、准确和低成本的检测方法用于常规监测高血压。 风险组（即，年轻、性活跃、多性伴）和监测对治疗的反应和复发。 因此，在健康检查期间用于唾液中HPV的非侵入性监测的POC测试可以具有显著的 影响和拯救生命。我们的目标是开发一种快速，易于使用的原型，用于检测HPV， 基于生物化学信号放大与CRISPR-Cas 12电化学的组合的唾液 读数和我们专有的坚固、准确和低成本的金叶电极。利用我们专有的金叶 电极沿着CRISPR-Cas 12测定用于靶核酸的灵敏和特异性电化学检测 酸性（NA）生物标志物，我们已经证明了用质粒DNA检测HPV 18以及在 病人标本我们提出这项建议的总体目标是开发一种可重复的工艺， 唾液，并开发用于HPV 16和HPV 18多重检测的原型电化学生物传感器 沿着参考基因以确保准确的测试结果。为了实现这一目标，我们将首先开发一个 我们的金箔电极用于检测HPV 16、HPV 18的空间多路复用的工作流程， 参考基因然后，我们将首先用质粒DNA，然后用唾液来验证这种多重测试 标本我们预计，将易于制造的空间多路复用电极与CRISPR- Cas 12 a用于检测特定的核酸序列将使我们的II期开发、调控 批准和商业化的一个点的需要的技术，可以规模和制造的准确， 易于使用和低成本的需求点检测，并减少HPV感染和OPSCC的负担。