Ultra-sensitive method for HIV p24 antigen detection

描述(由申请人提供)：这项建议的目标是发展超灵敏的ELISA(酶联免疫吸附试验)方法来定量检测HIV p24抗原。目前，p24抗原检测相对不敏感，只能检测到5-10pg/ml。即使病毒活跃复制，感染者的血清中也可能不存在这一数量的抗原。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，发展更灵敏的p24抗原检测具有重要意义。正是在这一背景下，我们提议开发一种高度敏感的p24抗原检测方法，该方法将比核糖核酸病毒载量检测便宜，并适用于发达国家和发展中国家，在发达国家和发展中国家，由于基础设施和实验室能力的限制，核酸检测无法进行。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检测准确性和功能性的所有变量；2)验证HIV-1 p24检测方法。所提出的分析方法具有灵敏度高、信噪比高、分析动态范围广、时间短、成本低等优点，证明了该方法具有显著的商业和科学潜力。公共卫生相关性：拟议的方法的开发将允许使用艾滋病毒p24抗原测试来监测艾滋病毒感染、血液筛查、急性感染的识别、协助诊断新生儿感染以及检测培养上清液中的抗原。与传统的检测方法相比，所开发的检测方法的性能将有显著的提高。成功完成这些研究将有利于公众健康，并为获得许可技术和制造商业产品的大量商业机会提供所需的所有技术。

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.