Promote HPV screening rate with a non-invasive HPV POC cartridge

 描述（由申请人提供）：HPV的持续感染导致癌症，并且可以预防-大约7900万美国妇女感染HPV，每年约有1400万新感染。2010年，有11，818名美国妇女被诊断患有宫颈癌，3939名宫颈癌死亡。HPV持续感染是导致宫颈癌发生的病理过程中的一个必要因素。宫颈癌是妇女中的一种主要致命恶性肿瘤，每年在全世界造成约275 000人死亡，其中大部分在发展中国家。宫颈癌如果在癌前阶段被发现并通过切除手术治疗，也是一种高度可预防的疾病。目前的HPV检测不筛查持续性HPV感染- HPV检测在识别HPV相关宫颈癌前病变或癌症风险增加的个体方面具有临床作用。目前，美国食品和药物管理局（FDA）批准了四种检测方法，用于检测13-14种高危HPV类型中任何一种的临床显著水平：（1）Qiagen Digene Hybrid capture（r）2（HC 2，Qiagen，盖瑟斯堡，MD），（2）Hologic-Genprobe Cervista（r）HPV（Hologics，贝德福德，MA），（3）Hologic-Genprobe APTIMA（r）HPV（Hologics，贝德福德，MA），和（4）Roche cobas（r）4800 HPV测试（Roche Molecular Systems，普莱森顿，CA）。但FDA批准的这些测试的适应症是高危HPV感染，而不是持续的高危HPV感染。研究表明，超过90%的新HPV感染，包括高危型，在两年内清除或无法检测，清除通常发生在感染后的前6个月。宫颈癌的主要特征之一是其从正常宫颈组织缓慢进展到组织中的癌前（或发育不良）变化，再到浸润性癌症。宫颈上皮内瘤变（CIN 3）进展为浸润性癌的平均时间估计为10至15年，并且有一小部分快速进展的宫颈癌在确诊阴性的巴氏试验后3年内被诊断出来。因此，大多数高危型HPV感染将在6个月内被人体免疫系统清除，只有一小部分高危型HPV感染会持续存在，并在10至15年后发展为浸润性宫颈癌。目前FDA批准的罗氏cobas HPV DNA检测的阳性hrHPV结果立即要求进行阴道镜检查，而大多数hrHPV感染不会持续。然而，在首次阳性hrHPV感染后6个月进行随访检测是困难的，因为目前的HPV检测不容易获得，不友好和侵入性。高危型HPV持续感染是宫颈癌前病变和浸润性宫颈癌最重要的危险因素。宫颈癌主要是未筛查或很少筛查的妇女的疾病。只有不到0.015%的HPV感染会导致浸润性宫颈癌，因此， 检测将检测持续的高危HPV感染，而不仅仅是高危HPV感染。不适和不便是后续检测确认持续感染的两个主要障碍。因此，对非侵入性样本（如尿液和唾液）进行自我HPV POC检测，沿着教育，可以显着提高HPV筛查率，与标准护理相比，女性一生中的一次HPV检测可以将癌症死亡率降低48%。这项SBIR工作的最终目标是通过展示安装有PocketLab的智能手机实现的非侵入性POC HPV检测来提高HPV筛查率。在接下来的II期研究中，HPV检测组将扩展到HPV 16/18 DNA，其他12种hrHPV DNA，E6/E7癌蛋白的库，系统控制和试剂输送将由手掌大小的PocketLab取代。这项拟议的SBIR I期研究将展示一种基于智能手机的HPV筛查设备，用于检测尿液、唾液和拭子等非侵入性标本中的hrHPV DNA和癌蛋白E6/E7。所有四种FDA批准的HPV DNA检测都是由训练有素的实验室人员使用的，他们精通在设备齐全的临床诊断实验室中进行实时PCR检测。这些测试需要使用复杂而昂贵的实验室设备，而这些设备在发展中国家并不普遍，并且需要几个人工处理步骤，这增加了污染的机会。所提出的HPV PocketLab检测盒将HPV扩增测定和免疫测定的样品处理、靶扩增和检测步骤集成到单个廉价的一次性检测盒中，该检测盒包含所有必需的冻干试剂，并且除了样品加载之外不需要手动步骤。

 DESCRIPTION (provided by applicant): Persistent infection by HPV causes cancers, and can be prevented - About 79 million American women are infected with HPV with about 14 million becoming newly infected each year. In 2010, there were 11,818 American women diagnosed with cervical cancer, and 3939 cervical cancer deaths. Persistent infection by HPV is a necessary factor in the pathologic process which may lead to cervical cancer development. Cervical cancer is a major fatal malignancy among women, causing about 275,000 deaths annually worldwide, mostly in developing countries. Cervical cancer is also a highly preventable disease if detected at its precancerous stages and treated by ablative procedures. The current HPV tests do not screen for persistent HPV infection - HPV testing has a clinical role in identifying individuals with an increased risk of an HPV-associated cervical precancer or cancer. Four tests are currently approved by the Food and Drug Administration (FDA) for detecting clinically significant levels of any of 13-14 high-risk HPV types: (1) the Qiagen Digene Hybrid capture(r) 2 (HC2, Qiagen, Gaithersburg, MD), (2) the Hologic-Genprobe Cervista(r) HPV (Hologics, Bedford, MA), (3) the Hologic-Genprobe APTIMA(r) HPV (Hologics, Bedford, MA), and (4) the Roche cobas(r) 4800 HPV test (Roche Molecular Systems, Pleasanton, CA). But the FDA approved indication of these tests is high-risk HPV infection, not persistent high risk HPV infection. Studies have shown that more than 90% of new HPV infections, including those with high-risk types, clear or become undetectable within two years, and clearance usually occurs in the first 6 months after infection. One of the key features of cervical cancer is its slow progression from normal cervical tissue, to precancerous (or dysplastic) changes in the tissue, to invasive cancer. The average time for progression of cervical intraepithelial neoplasia (CIN 3) to invasive cancer has been estimated to be 10 to 15 years, and there is a small subset of rapidly progressive cervical cancers which are diagnosed within 3 years of a confirmed negative Pap test. Hence, most high risk HPV infections will be cleared by body's immune system in 6 months, and only a smaller percentage of high risk HPV infections become persistent and develop into invasive cervical cancer after 10 to 15 years. A positive hrHPV result from the current FDA approved Roche's cobas HPV DNA test immediately calls for a colposcopy, while most hrHPV infections will not become persistent. However, it is difficult to conduct a follow-up test 6 months after the first positive hrHPV infection, because the current HPV tests are not easily accessible, unfriendly and invasive. Persistent infection with high-risk HPV is the most important risk factor for cervical cancer precursors and invasive cervical cancer. Cervical cancer is primarily a disease among unscreened or rarely screened women. Only less than 0.015% of HPV infection will lead to invasive cervical cancer, therefore a more clinically relevant screening test would detect persistent high-risk HPV infection, not just high-risk HPV infection. Discomfort and inconvenience are two major barriers of the follow up test to confirm persistent infection. Therefore, a self-administered HPV POC test on non-invasive samples such as urine and saliva along with education can significantly improve the HPV screening rate, and one HPV test in a woman's lifetime can decrease cancer mortality by 48% as compared to standard of care. The ultimate goal of this SBIR effort is to increase the HPV screening rate by demonstrating a non-invasive POC HPV test enabled by a smartphone mounted with a PocketLab. In the following Phase II study, the HPV test panel will be expanded to HPV 16/18 DNA, a pool of 12 other hrHPV DNA, E6/E7 oncoproteins, and the system control and reagent delivery will be replaced by a palm-sized PocketLab. This proposed SBIR Phase I study will demonstrate a smartphone-based HPV screening device to detect hrHPV DNA and oncoproteins E6/E7 from non-invasive specimens such as urine, saliva and swabs. All four FDA approved HPV DNA tests are intended for use by trained laboratory personnel who are proficient in performing real-time PCR assays in a well equipped clinical diagnostics laboratory. These tests require access to sophisticated and expensive lab equipment which is not widely available in developing countries, and several manual processing steps which increase the chances for contaminations. The proposed HPV PocketLab cartridge integrates the sample processing, target amplification, and detection steps of the HPV amplification assay and immunoassay into a single inexpensive, disposable cartridge that contains all necessary lyophilized reagents and requires no manual steps other than sample loading.