A Low-Cost Portable Multiplex Serum Molecular Test for HBV and HCV Detection

 描述（由申请人提供）：B肝炎（HB）和丙型肝炎（HC）分别是由血液传播的HB病毒（HBV）和HC病毒（HCV）引起的全球最广泛的两种感染。约有2.4亿人和1.5亿人分别患有慢性HB（CHB）和慢性HC（CHC），而每年约有780，000人和350，000 - 500，000人分别死于HB和HC的后果。在高度流行的国家，HBV和HCV的合并感染也很常见。HB和HC对医护人员和需要输血或器官移植的患者也是重要的危害，因为CHB和CHC无症状，患者和血液/组织供体可能在不知情的情况下患有HB或HC，对医护人员和需要输血或组织移植的患者构成风险。目前，HB和HC是用两套单独的测试来诊断的，这两套测试可以在最初感染后几周或几个月内检测到HB和HC。定量PCR（qPCR）可以在感染的第一周内检测HB和HC，但它们价格昂贵，不适合在HB和HC流行的低资源国家使用。他们也是两个测试，加起来的成本。该研究的目的是开发一种低成本、快速、易用、多重和准确的血清检测方法，在一次检测中同时检测HBV DNA和HCV RNA，以在低资源HB和HC流行国家的护理点诊断HB和HC，并筛选血液和器官供应。（PbMg_（1/3）Nb_（2/3）O_3）_（0.65）（PbTiO_3）_（0.35）（PMN-PT）压电传感器（PEPS）是一种利用相对共振频移（Δ f/f）检测靶分子与PEPS表面受体结合的新型传感器。PMN-PT PEPS的独特之处在于，由于PMN-PT层内独特的偏振切换机制，检测的λ/f/f增强了1000倍。因此，PEPS在DNA检测中表现出类似于PCR的灵敏度，但不需要DNA提取和DNA扩增。Lenima Field Diagnostics将开发3-PEPS阵列，其中一个PEPS检测HBV DNA，第二个PEPS检测HCV RNA，第三个PEPS作为阴性对照。初步结果表明，PEPS HBV DNA检测血清中加标HBV的分析灵敏度为100个病毒/ml，与qPCR的灵敏度115个病毒/ml相似。使用类似的策略，PEPS在40名患者粪便中艰难梭菌毒素B基因检测的单独研究中证明了95%的灵敏度和95%的特异性。预计我们将成功开发多重PEPS HBV和HCV检测，以实现灵敏度>90%和特异性>90%的里程碑。在第二阶段，我们将开发临床原型，包括便携式检测系统和一次性阵列PEPS，用于FDA批准的患者样本检测。

 DESCRIPTION (provided by applicant): Hepatitis B (HB) and hepatitis C (HC) are two of the most widespread infections worldwide caused by the blood-borne HB virus (HBV) and HC virus (HCV), respectively. About 240 million and 150 million people suffer from chronic HB (CHB) and chronic HC (CHC), respectively while about 780,000 and 350,000-500,000 people die each year due to the consequence of HB and HC, respectively. In highly endemic countries, co-infection of HBV and HCV is also common. HB and HC are also important hazards for healthcare workers and for patients needing transfusion or organ transplant as CHB and CHC are asymptomatic and patients and blood/tissue donors may have HB or HC without knowing it, posing risks to healthcare workers and patients needing transfusion or tissue transplant. Currently HB and HC are diagnosed with two separate sets of tests which can detect HB and HC only weeks or months after the initial infection. Quantitative PCR (qPCR) could detect both HB and HC within the first week of infection but they are expensive not suitable for use in low-resource countries where HB and HC are endemic. They are also two tests, adding up the cost. The goal of the proposed study is to develop a low-cost, rapid, easy-to-use, multiplex, and accurate serum test to detect both HBV DNA and HCV RNA in one test to diagnose HB and HC at point of care, in low-resource HB and HC endemic countries, and to screen for blood and organ supplies. (PbMg1/3Nb2/3O3)0.65(PbTiO3)0.35 (PMN-PT) piezoelectric plate sensor (PEPS) is a new sensor that uses relative resonance frequency shift, f/f to detect binding of a target to the receptor on the PEPS surface. Unique to PMN-PT PEPS is that the detection f/f is enhanced by 1000 times due to the unique polarization switching mechanism within the PMN- PT layer. As a result, PEPS has demonstrated sensitivity similar to PCR in DNA detection but without the need of DNA extraction and DNA amplification. Lenima Field Diagnostics will develop a 3-PEPS array with one PEPS detecting HBV DNA, a second PEPS detecting HCV RNA, a third PEPS serving as a negative control. Preliminary results indicate 100 viruses/ml analytical sensitivity fr PEPS HBV DNA detection of HBV spiked in serum as similar to the 115 viruses/ml sensitivity of qPCR. Using a similar strategy, PEPS has demonstrated 95% sensitivity and 95% specificity in a separate study of Clostridium difficile toxin B gene detection in 40 patients' stools. It is expected that we will successfully develop the multiplexed PEPS HBV and HCV test to achieve a milestone of >90% sensitivity and >90% specificity. In phase II, we will develop the clinical prototype that includes a portable detection system and disposable array PEPS for patient sample tests towards FDA approval.