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

 描述(申请人提供)：HPV持续感染会导致癌症，并且是可以预防的-美国约有7900万女性感染HPV，每年约有1400万人新感染。2010年，有11818名美国女性被诊断患有宫颈癌，3939人死于宫颈癌。HPV持续感染是宫颈癌发生发展过程中的必要因素。宫颈癌是女性的一种主要致命恶性肿瘤，每年导致全球约275,000人死亡，其中大部分发生在发展中国家。宫颈癌也是一种高度可预防的疾病，如果在癌前阶段被发现并通过消融术进行治疗。目前的HPV检测并不筛查持续的HPV感染-HPV检测在临床上起到了识别HPV相关宫颈癌前病变或癌症风险增加的个人的作用。食品和药物管理局(FDA)目前批准了四种检测13-14种高危HPV类型中任何一种的临床显著水平的检测方法：(1)Qiagen Digene杂交捕获(R)2(HC2，Qiagen，Gaithersburg，MD)，(2)Hology-GenProbe Cervista(R)HPV(Hologics，Bedford，MA)，(3)Hology-GenProbe APTIMA(Hologics，Bedford，MA)，和(4)Roche Cobas(R)4800 HPV检测(罗氏分子系统公司，加利福尼亚州普莱森顿)。但FDA批准的这些检测的适应症是高危HPV感染，而不是持续性的高危HPV感染。研究表明，超过90%的新HPV感染，包括高危类型的HPV感染，在两年内消失或无法检测到，并且通常在感染后的前6个月清除。宫颈癌的主要特征之一是其从正常宫颈组织到癌前(或发育不良)组织的缓慢进展，再到浸润性癌。宫颈上皮内瘤变(CIN 3)发展为浸润性癌症的平均时间估计为10至15年，有一小部分快速进展的宫颈癌在巴氏试验阴性后3年内被诊断出来。因此，大多数高危HPV感染将在6个月内被人体免疫系统清除，只有较小比例的高危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批准的罗氏眼镜蛇HPV DNA检测结果呈阳性，立即需要进行阴道镜检查，而大多数hrHPV感染不会持续存在。然而，很难在首次hrHPV感染阳性后6个月进行后续检测，因为目前的HPV检测不容易获得，不友好，而且具有侵入性。高危型HPV持续感染是宫颈癌先兆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一种疾病。只有不到0.015%的hpv感染会导致浸润性宫颈癌，因此更具临床相关性的筛查。 检测将检测持续的高危HPV感染，而不仅仅是高危HPV感染。不适和不便是后续测试确认持续感染的两大障碍。因此，对尿液和唾液等非侵入性样本进行自我管理的HPV POC检测结合教育可以显著提高HPV筛查率，与标准护理相比，女性一生中进行一次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一种由安装了PocketLab的智能手机实现的非侵入性POC HPV检测来提高HPV筛查率。在接下来的第二阶段研究中，HPV测试板将扩大到HPV16/18DNA，12个其他hrHPV DNA，E6/E7癌蛋白的池，系统控制和试剂输送将被一个手掌大小的PocketLab取代。这项拟议的SBIR第一阶段研究将展示一种基于智能手机的HPV筛查设备，用于从尿液、唾液和拭子等非侵入性样本中检测hrHPV DNA和癌蛋白E6/E7。FDA批准的所有四种HPV DNA测试都旨在供训练有素的实验室人员使用，他们熟练地在设备齐全的临床诊断实验室进行实时PCR分析。这些检测需要获得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存在，而且需要几个人工处理步骤，这增加了污染的可能性。建议的HPV PocketLab试剂盒将HPV扩增分析和免疫分析的样品处理、靶标扩增和检测步骤整合到一个廉价的一次性试剂盒中，该试剂盒包含所有必要的冻干试剂，除样本加载外不需要其他手动步骤。

 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.