Enzyme-linked immunoassay to detect
Indirect ELISAs to detect exposure to Felis catus gammaherpesvirus 1 (FcaGHV1) in domestic cats. These ELISAs detect feline serum antibodies to ORF52 and ORF38 of FcaGHV1. The ELISA assays are sensitive, specific, and adaptable for scale up use in high throughput diagnostics.
1. A method of detecting antibodies to Felis catus gammaherpesvirus 1 (FcaGHV1) in a test sample obtained from a feline comprising the steps of:
obtaining a test sample, wherein the test sample is blood, serum or plasma sample; and
performing an immunoassay comprising:
contacting the test sample with full-length recombinant FcaGHV1 ORF38 and ORF52 polypeptides; and
detecting binding between antibodies to FcaGHV1 in the test sample and at least one of the full-length recombinant FcaGHV1 polypeptides, whereby the presence of binding between antibodies to FcaGHV1 in the sample and one of the recombinant FcaGHV1 polypeptides is indicative of antibodies to FcaGHV1 in the test sample.
2. The method according to claim 1 , wherein the test sample obtained from a feline is a test sample from a domestic cat.
3. The method according to claim 1 wherein the immunoassay is an IFA, a western blot assay or an ELISA.
4. The method according to claim 1 wherein the immunoassay is an indirect ELISA.
5. The method of detecting antibodies to FcaGHV1 in a test sample according to claim 1 , wherein the test sample is blood, further comprising the step of performing qPCR on the blood sample to screen for FcaGHV1 nucleic acid.
6. The method of detecting antibodies to FcaGHV1 in a test sample according to claim 1 further comprising:
contacting the test sample with additional full-length recombinant FcaGHV1 polypeptides selected from the group consisting of ORF17.5, ORF65, LANA, and combinations thereof; and
detecting binding between antibodies to FcaGHV1 in the test sample and at least one of the additional recombinant FcaGHV1 polypeptides.
7. A method of detecting antibodies to Felts catus gammaherpesvirus 1 (FcaGHV1) in a test sample obtained from a feline comprising the steps of:
obtaining the test sample, wherein the test sample is a blood, serum or plasma sample;
performing an immunoassay comprising:
contacting the test sample with full-length recombinant FcaGHV1 ORF38 and ORF52 polypeptides; and
detecting binding between antibodies to FcaGHV1 in the test sample and at least one of the full-length recombinant FcaGHV1
polypeptides, whereby the presence of binding between antibodies to FcaGHV1 in the sample and one of the recombinant FcaGHV1 polypeptides is indicative of antibodies to FcaGHV1 in the test sample; and
performing qPCR on the sample, wherein the sample is blood, to screen for FcaGHV1 nucleic acid.