Serological screening for HHV-8 infection using antigen mixtures
Inventors
Burbelo, Peter D. • Kovacs, Joseph • Iadarola, Michael J.
Assignees
US Department of Health and Human Services
Publication Number
US-8951723-B2
Publication Date
2015-02-10
Expiration Date
2030-02-12
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Abstract
The invention provides compositions, methods, and kits for the diagnosis or detection of active, latent, or prior infection with human herpesvirus 8 in a subject sample.
Core Innovation
The invention provides compositions, methods, and kits for the diagnosis or detection of active, latent, or prior infection with human herpesvirus 8 (HHV-8) in a subject sample. The methods include contacting the subject sample with at least an antigenic fragment of an HHV-8 v-cyclin polypeptide and detecting specific binding of an antibody in the sample to indicate HHV-8 infection.
The invention further offers assays utilizing multiple antigenic fragments from HHV-8 polypeptides including K8.1, ORF65, and LANA to improve detection sensitivity and specificity. These antigenic fragments can be present in a single reaction mixture or tested simultaneously or sequentially, enabling stratification of subjects based on antibody profiles.
Claims Coverage
The patent contains multiple independent claims focusing on methods of detecting HHV-8 infection using antigenic fragments of HHV-8 proteins, primarily v-cyclin, alone or in combination with other HHV-8 antigens.
Detection of HHV-8 infection using HHV-8 v-cyclin antigenic fragment linked to a reporter polypeptide
A method of detecting active, latent, or prior HHV-8 infection by contacting a subject sample with at least an antigenic fragment of HHV-8 v-cyclin polypeptide covalently linked to a reporter polypeptide, detecting specific binding of antibodies in the sample to this fragment, wherein this binding indicates HHV-8 infection.
Use of multiple HHV-8 antigenic fragments for enhanced detection
Methods comprising contacting the sample with antigenic fragments of one or more of HHV-8 K8.1, ORF65, and LANA polypeptides in addition to the v-cyclin fragment to detect antibodies binding to these fragments for diagnosis.
Specific sequences of antigenic fragments
Utilization of defined amino acid sequence ranges as antigenic fragments for K8.1 (e.g., residues 25-228, 143-228, etc.), ORF65 (e.g., residues 2-170), and LANA (various fragments such as 274-925, 6-286, and others) in the detection methods.
Simultaneous detection using antigen mixtures
Performing detection by contacting a sample with two, three, or four antigenic fragments from different HHV-8 polypeptides in a single reaction mixture to improve assay throughput and sensitivity.
Identification and stratification of HHV-8 infection states
In a subject diagnosed with HHV-8 infection, comparing antibody amounts bound to antigenic fragments of v-cyclin or LANA relative to ORF65 to distinguish between lytic and latent HHV-8 infections.
The independent claims cover methods of detecting HHV-8 infection using antigenic fragments of v-cyclin alone or combined with antigenic fragments of K8.1, ORF65, and LANA, employing specific polypeptide sequences and mixtures in a single assay, and further include stratification of infection status based on differential antibody binding.
Stated Advantages
Provides highly specific and highly sensitive methods for detection of HHV-8 infection, achieving sensitivity and specificity of at least 95%, and in some embodiments up to 100%.
Enables efficient high-throughput screening using antigen mixtures, simplifying data collection and analysis.
Offers improved diagnostic performance compared to existing assays like ELISA, including detection of infections that ELISA might miss.
Allows stratification of subjects into latent or lytic infection states based on antibody profiles to different HHV-8 antigens.
Suitable for screening blood supplies and donated tissues or organs to ensure safety from HHV-8 infection.
Documented Applications
Diagnosis or detection of active, latent, or prior HHV-8 infection in individual subjects using serological assays.
Screening blood supplies after donor collection for HHV-8 infection to ensure safety in transfusion.
Testing tissues and organs from donors, including cadaver donors, to verify absence of HHV-8 infection prior to transplantation.
Monitoring HHV-8 associated cancers such as Kaposi sarcoma, including potential use of anti-v-cyclin antibodies in cancer detection and monitoring.
Monitoring AIDS disease progression or regression related to susceptibility to HHV-8 infection.
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