There are many pathogenic organisms that cause disease in the oral cavity or that can be detected there. Among these are three important viruses, human immunodeficiency virus (HIV), human papilloma virus (HPV) and herpes simplex virus (HSV). HIV infection and disease is treatable but is costly and is still associated with considerable morbidity and mortality. HPV is the primary cause of oropharyngeal cancer and HSV causes common oral lesions. All of these pathogens can be spread by contact with saliva. This is not a common route of transmission for HIV, but it is nevertheless a concern since HIV causes AIDS which is a fatal disease. It would therefore be desirable to have a simple, cost-effective point-of-care (POC) device for the detection of infection by HIV, HPV and HSV. We propose to use a protein microarray to detect HIV by antigen capture and to detect antibodies to each virus based on binding to viral proteins in the microarray. Antigen Discovery Incorporated (ADi) has considerable experience in the development, production and marketing of protein microarrays. We now sell seventeen reactive protein antigen microarrays from a variety of pathogenic viruses, bacteria and protozoa including an HIV-1 array with proteins from the five most common subtypes. Moreover, ADi has already isolated all the genes from eleven serotypes of HPV, HSV types 1 and 2 and inserted them into an expression vector for production of proteins. In order to develop a useful POC device for the detection of HIV as well as antibodies against HIV, HPV and HSV in saliva, we have the following specific aims for the first phase of this project: 1. Create a proteomic microarray containing all proteins from all common types of HIV-1, HPV and HSV. Later we will expand the microarray to include other oral pathogens 2. Develop a protocol for sensitive and specific screening for antibodies to oral pathogens in saliva using a proteomic microarray. In Phase II a microfluidic device will be developed to automate the assay procedure. 3. Develop a sensitive and specific microarray-based antigen-capture detection assay for HIV. In Phase II similar assays will be developed for HPV, HSV and other oral pathogens and the assay procedure will be automated using a microfluidic device.
This project will create a prototype point-of-care device for detection of HIV and antibodies against HIV, HPV and HSV in saliva. We will develop a simple protocol for use of the device that will ensure sensitive, specific detection of oral pathogens and antibodies against them in saliva.
