Digital LAMP as a point of care assay for HPV detection

数字 LAMP 作为 HPV 检测的护理点检测

• 批准号: 9071862
• 负责人: Daniel T Chiu
• 金额: $ 49.09万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-04-01 至 2019-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9071862
• 关键词: Archives; Biological Assay; Cervical; Cervical Cancer Screening; Characteristics; Clinic; Clinic Visits; Clinical; Cytology Histology; Detection; Developed Countries; Developing Countries; Diagnosis; Equipment; Genital Human Papilloma Virus Infection; Genital system; Genotype; HPV-High Risk; High Risk Woman; Histologic; Home environment; Human Papillomavirus; Human papilloma virus infection; Human papillomavirus 16; Individual; Infection; Lesion; Malignant neoplasm of cervix uteri; Mediating; Methods; Microfluidic Microchips; Nucleic Acids; Oncogenic; Pap smear; Patients; Performance; Population; Premalignant; Preparation; Reporting; Resources; Rural; Sampling; Sensitivity and Specificity; Specificity; Swab; Testing; Uninsured; United States Food and Drug Administration; Urine; Vagina; Viral Load result; Visit; Woman; aqueous; base; digital; flexibility; follow-up; high risk; improved; infection related cancer; innovative technologies; instrument; mortality; nanolitre; novel; point of care; public health relevance; rapid diagnosis; screening; success; uptake

 DESCRIPTION (provided by applicant): We propose to develop a digital loop-mediated-isothermal-amplification (dLAMP), point-of-care (POC), instrument-free, assay for genital human papillomavirus (HPV) testing that can be used as a primary screening method for cervical cancer. Recently, the US Food & Drug Administration approved the Roche Cobas HPV test for stand-alone primary cervical cancer screening. This test, however, requires a sophisticated lab setting and equipment, and thus is not feasible for use as a POC assay, certainly not in a self-test setting, but also not in rural clinical or in resource-poor settings. We have recently developed a novel digital nucleic acid amplification platform based on a self-digitization (SD) microfluidic chip, which partitions an aqueous sample into tens of thousands of nanoliter volumes. Using this SD platform, we have also developed and reported the first digital LAMP assay. Based on these two innovative technologies, we propose to develop a POC HPV detection assay fulfilling the following criteria: (1) It should be as sensitive and specific as existing FDA-approved HPV testing assays for detecting precancerous lesions; (2) It should be instrument-free and require minimal sample preparation, thus offering the flexibility to be done by the patient herself at home; (3) It should provide viral load information, which can be used to improve the specificity of HPV testing and reduce over-treatment; (4) It should be flexible enough to incorporate testing for HPV integration and additional markers to improve specificity; and (5) It should be robust, fast, and affordable. We see application of our proposed POC assay in three different settings: (1) In developed countries, the POC assay can be performed at home and used for primary screening outside the clinical setting, thus maximizing the uptake of screening; (2) In developed countries, the POC assay can be used in the clinic for screening underserved and uninsured populations, allowing for rapid diagnosis and same-visit treatment, thus reducing patient loss to follow-up; (3) In developing countries, the POC assay will allow the clinicians to screen and treat patients at the same clinic visit, again reducing loss of high-risk women through follow-up. Thus, our proposed test will have significant impact on the success of cervical cancer screening in both developed and developing countries.

 描述（由申请人提供）：我们建议开发一种数字环介导等温扩增 (dLAMP)、即时护理 (POC)、无仪器、可用于生殖器人乳头瘤病毒 (HPV) 检测的检测方法最近，美国食品和药物管理局批准了罗氏 Cobas HPV 检测用于独立的宫颈癌初级筛查，但该检测需要复杂的实验室环境和设备。用作 POC 测定是不可行的，当然在自检环境中也不可行，而且在农村临床或资源匮乏的环境中也不可行。我们最近开发了一种基于自我数字化的新型数字核酸扩增平台（ SD）微流控芯片，可将水样分成数万纳升体积，我们还开发并报告了第一个基于这两项创新技术的数字 LAMP 检测。化验填充符合以下标准：（1）它应该与 FDA 批准的现有用于检测癌前病变的 HPV 检测方法一样敏感和特异；（2）它应该无需仪器，并且需要最少的样品制备，从而提供了灵活性； (3) 应提供病毒载量信息，可用于提高 HPV 检测的特异性并减少过度治疗； (4) 应足够灵活，可纳入 HPV 整合检测和其他标记物；提高特异性；以及（5）它应该是稳健、快速且经济实惠的，我们所提出的 POC 检测可在三种不同的环境中应用：（1）在发达国家，POC 检测可以在家中进行，并用于临床环境之外的初步筛查，从而最大限度地发挥作用。 (2) 在发达国家，POC 可用于诊所对服务不足和未投保人群进行筛查，从而实现快速诊断和同诊治疗，从而减少患者失访；在发展中国家，POC 检测将允许忠实患者在同一诊所就诊时进行筛查和治疗，再次通过随访减少高危女性的损失。因此，我们提出的测试将对发达国家和发展中国家宫颈癌筛查的成功产生重大影响。