Compositions and methods for screening for lyme disease
Inventors
Burbelo, Peter D. • Marques, Adriana • Iadarola, Michael J.
Assignees
US Department of Health and Human Services
Publication Number
US-9310367-B2
Publication Date
2016-04-12
Expiration Date
2031-03-10
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Abstract
The invention provides compositions, methods, and kits for the diagnosis or detection of infection by a pathogen that causes Lyme disease in a subject.
Core Innovation
The invention provides compositions, methods, and kits for the diagnosis or detection of infection by a Borrelia species pathogen that causes Lyme disease in a subject. The compositions comprise isolated peptides having amino acid sequences identical or substantially identical to specific sequences (SEQ ID NOs: 1-9), derived from Borrelia burgdorferi, Borrelia afzelii, Borrelia garinii, and/or Borrelia valaisiana. These peptides, alone or joined by covalent linkages such as peptide bonds or peptide linkers, can be used in immunoassays to detect antibodies in subject samples indicative of infection. The methods include contacting a sample with these peptide compositions and detecting binding to antibodies, which can be used for diagnosis, monitoring therapeutic treatment response, and selecting treatment regimens.
The problem being addressed is the difficulty in sensitive and specific diagnosis of Lyme disease, which is caused by Borrelia species and can present with variable symptoms. Current diagnostic methods such as culturing Borrelia bacteria are difficult, and serological tests including ELISA and Western blot have limitations in sensitivity, specificity, and throughput. The difficulty is compounded by the need for assays that can detect antibodies early in infection and with a broad dynamic range to monitor treatment response effectively. There is a particular need for improved, high-throughput, and reliable immunoassays for Lyme disease diagnosis and monitoring.
The invention introduces novel peptide antigens including a synthetic antigen called VOVO, which contains multiple immunoreactive peptide fragments derived from the VlsE and OspC proteins of Borrelia species. These antigenic peptides demonstrate superior sensitivity and specificity compared to existing assays (e.g., C6 ELISA) and allow detection of anti-Lyme antibodies without the need for serum dilution, offering a wide dynamic range. The approach includes the use of luciferase immunoprecipitation systems (LIPS) that employ recombinant fusion proteins of these peptides with reporter enzymes to quantitatively measure antibody titers in patient samples. The invention thus overcomes disadvantages of traditional assays by providing a sensitive, specific, and high-throughput platform for Lyme disease diagnosis and monitoring.
Claims Coverage
The claims include one independent claim focusing on a diagnostic method for Borrelia sp. infection and several dependent claims directed to specific features of the method.
Method of diagnosing Borrelia sp. infection by antibody binding
A method including obtaining a sample from a subject, contacting the sample with a composition comprising an isolated peptide containing the amino acid sequence SEQ ID NO: 1, and detecting binding of that peptide to an antibody in the sample as indicative of infection by Borrelia sp. selected from B. burgdorferi, B. garinii, and B. afzelii.
Isolation of antibody-peptide complex
The method further includes isolating the antibody-peptide complex from the sample as part of the detection process.
Use of isolated peptide consisting of SEQ ID NO: 1
The composition used in the method can comprise an isolated peptide consisting exactly of the amino acid sequence SEQ ID NO: 1.
Use of reporter polypeptide linked to peptide
The composition can comprise a reporter polypeptide covalently linked to the isolated peptide to facilitate detection.
The claims cover a diagnostic method using specific isolated peptides derived from Borrelia sequences for detecting infection by binding antibodies in subject samples, encompassing variations such as use of exactly defined peptides, linkage to reporter polypeptides, and isolation of antibody-peptide complexes, specifically targeting infections by Borrelia burgdorferi, Borrelia garinii, and Borrelia afzelii.
Stated Advantages
High sensitivity and specificity for detection of Lyme disease antibodies compared to existing assays such as C6 ELISA.
Ability to detect antibodies without serum dilution, providing a broad dynamic range of detection.
Suitability for high-throughput screening due to the use of recombinant peptide fusion proteins and luminescent immunoprecipitation assays.
Improved accuracy and practicality for diagnosing early infection and monitoring therapeutic treatment response.
Documented Applications
Diagnosis of infection by Borrelia sp. in subjects using peptide-based immunoassays.
Monitoring therapeutic treatment response in subjects with Borrelia sp. infection by assessing antibody binding to the peptide compositions.
Selecting treatment regimens for subjects based on susceptibility indicated by antibody binding to peptides.
Screening donated blood, tissues, or organs for Borrelia sp. infection prior to transplantation or transfusion.
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