Digital LAMP as a point of care assay for HPV detection

 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）、床旁（POC）、免仪器检测生殖器人乳头瘤病毒（HPV）检测方法，可用作宫颈癌的初步筛查方法。最近，美国食品和药物管理局批准罗氏Cobas HPV检测用于独立的原发性宫颈癌筛查。然而，该测试需要复杂的实验室设置和设备，因此不适合用作POC测定，当然不能用于自测设置，也不能用于农村临床或资源贫乏的设置。 我们最近开发了一种基于自数字化（SD）微流控芯片的新型数字核酸扩增平台，该芯片将水性样品分成数万纳升体积。使用这个SD平台，我们还开发并报道了第一个数字LAMP检测。基于这两项创新技术，我们建议开发一种符合以下标准的POC HPV检测方法：（1）它应该与现有FDA批准的HPV检测方法一样敏感和特异，用于检测癌前病变;（2）它应该是无仪器的，需要最少的样品制备，因此提供了患者自己在家完成的灵活性;（3）它应该提供病毒载量信息，可用于提高HPV检测的特异性并减少过度治疗;（4）它应该足够灵活，可以将HPV整合检测和其他标志物结合起来，以提高特异性;（5）它应该稳健，快速，负担得起。 我们看到我们提出的POC测定法在三种不同环境中的应用：（1）在发达国家，POC测定法可以在家里进行，并用于临床环境之外的初级筛查，从而最大化筛查的吸收;（2）在发达国家，POC检测可用于临床筛查服务不足和无保险的人群，实现快速诊断和同诊治疗，因此减少了患者的随访损失;（3）在发展中国家，POC测定将允许临床医生在同一次诊所就诊时筛查和治疗患者，再次减少了高风险妇女通过随访的损失。 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。

 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.