Ultra-sensitive method for HIV p24 antigen detection

描述（由申请人提供）：本提案的目的是开发用于定量HIV p24抗原的超灵敏ELISA（酶联免疫吸附试验）方法。目前，p24抗原检测相对不敏感，只能检测到5- 10pg /ml。这种数量的抗原可能不存在于感染者的血清中，即使病毒正在积极复制。事实上，只有大约50-60%的艾滋病患者、30-40%的ARC患者和10%的无症状患者会有可检测到的p24抗原血症。因此，开发更灵敏的p24抗原检测方法具有重要意义。正是在这种背景下，我们建议开发一种高度敏感的p24抗原测定方法，这种方法将比RNA病毒载量检测便宜，适合在基础设施和实验室能力的限制禁止进行核酸检测的发达国家和发展中国家使用。第一阶段和第二阶段的具体目标部分包括：1)分析和优化影响p24测试准确性和功能的所有变量；2)验证HIV-1 p24检测方法。该方法具有灵敏度高、信噪比大、动态范围广、分析时间短、成本低等优点，具有显著的商业和科学潜力。公共卫生相关性：拟议方法的发展将允许使用艾滋病毒p24抗原检测来监测艾滋病毒感染、血液筛查、识别急性感染、协助诊断新生儿感染和检测培养上清中的抗原。与传统检测方法相比，开发的检测方法的性能将得到显著改善。这些研究的成功完成将有利于公共卫生，并提供所需的所有技术，以获得技术许可和制造商业产品的大量商业机会。

DESCRIPTION (provided by applicant): The objective of this proposal is to develop ultra-sensitive ELISA (Enzyme-Linked ImmunoSorbent Assay) methods for quantification of HIV p24 antigen. At present, the p24 antigen test is relatively insensitive, being able to detect only 5- 10 pg/ml. This quantity of antigen may not be present in the serum of infected individuals, even when the virus is actively replicating. In fact, only about 50-60% of AIDS patients, 30-40% of ARC patients, and 10% of asymptomatic patients will have p24 antigenemia that is detectable. Therefore, the development of more sensitive p24 antigen tests is of great importance. It is in this context that we are proposing to develop a highly sensitive p24 antigen assays that will be less expensive than RNA viral load tests and appropriate for use both in developed and developing countries in which the limitations of infrastructure and laboratory capability prohibit nucleic acid testing. The specific aims of the Phase I and Phase II, in part, include 1) analyzing and optimizing of all variables affecting the accuracy and functionality of the p24 tests; and 2) validation of the HIV-1 p24 detection assay. Advantages of the proposed assays such as the increased sensitivity, signal-to- noise ratio, and dynamic range, reduced time and low cost of the analysis substantiates our belief that the proposed amplification method has a prominent commercial and scientific potential. PUBLIC HEALTH RELEVANCE: The development of the proposed method will allow using HIV p24 antigen tests for monitoring HIV infection, blood screening, identification of acute infection, to assist in the diagnosis of infection in the newborn, and in detecting antigen in supernatants from cultures. The performance of the developed tests will be improved significantly as compared with that for conventional assays. Successful completion of these studies will be beneficial for public health, and make available all of the technology needed for a substantial business opportunity to license the technology and manufacture commercial products.