초록 ▼

This invention provides methods and compositions for testing for pregnancy and non-pregnancy in ungulates and non-hoofed ruminates. The tests provided by this invention are useful during a time that coincides with the estrus cycle during which breeding occurs or the first estrus cycle after breeding

This invention provides methods and compositions for testing for pregnancy and non-pregnancy in ungulates and non-hoofed ruminates. The tests provided by this invention are useful during a time that coincides with the estrus cycle during which breeding occurs or the first estrus cycle after breeding of a non-pregnant animal. The tests provided by this invention are useful in estrus and ovulation synchronization programs, with pregnancy testing useful at a time allowing for resynchronization of non-pregnant animals within the first estrus cycle. The tests provided by this invention assay for the presence, absence, or level of a selected IFN-τ-induced protein in a sample from a female animal. The tests of this invention are useful for testing cells, blood, plasma, serum, cells, milk, nasal secretions, ocular secretions, vaginal secretions, urine, and saliva samples. The tests provided by this invention are immunoassays. Polyclonal and monoclonal antibodies useful in such tests, as well as methods of making such antibodies and hybridoma cell lines, are provided. Devices for performing such tests, methods of using such devices, and methods of making such devices are provided. Kits containing such devices are also provided. This invention provides a method for determining readiness for breeding. This invention provides a method for resynchronizing breeding with breeding cycle times of one estrus cycle or shorter. This invention also provides a method for breeding by forcing estrus and artificial insemination by appointment.

대표청구항 ▼

We claim: 1. A method of detecting pregnancy in a female sheep or cow comprising detecting interferon-stimulated gene 17 (ISG17) in a test sample of blood or serum from a female sheep or cow, said method comprising: (a) providing an immunoassay test device comprising: i. at least one absorbent supp

We claim: 1. A method of detecting pregnancy in a female sheep or cow comprising detecting interferon-stimulated gene 17 (ISG17) in a test sample of blood or serum from a female sheep or cow, said method comprising: (a) providing an immunoassay test device comprising: i. at least one absorbent support; ii. an antibody specific to said ISG17 on a said absorbent support; iii a flow control component on a said absorbent support; iv. means for detecting binding of said antibody with said ISG17; and v. means for comparing the binding level of ISG17 in the test sample with a background binding level of ISG17 in a control sample from a non-pregnant animal of the same species; (b) flowing the sample into the immunoassay test device into contact with the antibody; (c) detecting a binding level of said antibody with said ISG17 in the test sample greater than that of the binding level of said antibody with ISG17 in a control sample from a non-pregnant animal of the same species, said greater binding level of said antibody with ISG17 in the test sample being indicative of pregnancy; and wherein said method does not comprise a cell-lysing step. 2. The method of claim 1 comprising collecting said sample from said sheep or cow and detecting said binding outside a laboratory. 3. The method of claim 1 wherein said test sample is obtained on a day selected from the group consisting of Day 11, Day 32, and intervening days after breeding. 4. The method of claim 3 wherein said breeding comprises performing artificial insemination. 5. The method of claim 3 wherein said breeding is preceded by forced estrus. 6. The method of claim 1 wherein said test sample is obtained on a day selected from the group consisting of Day 15, Day 25, and intervening days after breeding. 7. The method of claim 1 wherein said test sample is obtained on a day selected from the group consisting of Day 18, Day 22, and intervening days after breeding. 8. The method of claim 1 wherein said sample is obtained on a day selected from Day 18 or Day 19 after breeding. 9. The method of claim 1 wherein said test sample is obtained on Day 18 after breeding. 10. The method of claim 1 wherein said test sample is obtained on Day 19 after breeding. 11. The method of claim 1 wherein said sheep or cow is not infected with a virus. 12. The method of claim 1 wherein said test sample is blood. 13. The method of claim 12 also comprising filtering the blood sample to obtain serum containing said ISG17. 14. The method of claim 1 performed outside a laboratory. 15. The method of claim 1 wherein said antibody is selected from the group consisting of: monoclonal and polyclonal antibodies generated with and capable of specific binding to an amino acid sequence selected from the group consisting of QRLAHLDSREVLQE (SEQ ID NO: 1), CQRLAHLDSREVLQE (SEQ ID NO: 2), TVAELKQQVCQKERVQ (SEQ ID NO: 3), CTVAELKQQVCQKERVQ (SEQ ID NO: 4), WLSFEGRPMDDEHPLE (SEQ ID NO: 5), and CWLSFEGRPMDDEHPLE (SEQ ID NO: 6); and monoclonal and polyclonal antibodies generated with other complete or partial amino acid sequences from mammalian ISG17. 16. The method of claim 1 comprising comparing binding of said antibody with ISG17 in said test sample with binding of said antibody in a second control sample containing normal background levels of said ISG17. 17. The method of claim 16 also comprising binding a second antibody to said ISG17 in said test sample. 18. The method of claim 1 wherein said antibody specific to ISG17 is 5E9, wherein a hybridoma that produces said antibody is deposited as ATCC Deposit No. PTA-7960. 19. The method of claim 1 also comprising binding a second antibody to ISG17 to said ISG17 in said test sample. 20. The method of claim 19 wherein said antibody and said second antibody bind to substantially distinct epitopes. 21. A method of breeding of a female sheep or cow comprising: (1) detecting non-pregnancy in the sheep or cow by a method comprising: (a) on a day that is after a first breeding of said sheep or cow, wherein said sheep or cow was bred during estrus, said day being before estrus of said sheep or cow is next expected, obtaining a sample of blood or serum from said sheep or cow; (b) providing an immunoassay device comprising: i. at least one absorbent support; ii. an antibody specific to said ISG17 on a said absorbent support; iii a flow control component on a said absorbent support; iv. means for detecting binding of said antibody with said ISG17; and v. means for comparing the binding level of ISG17 in the test sample with a background binding level of ISG17 in a control sample from a non-pregnant animal, a pregnant animal or a recently-aborted animal of the same species; (c) flowing the sample into the immunoassay test device into contact with the antibody; (d) detecting a binding level of said antibody with said ISG17 in the test sample not greater than that of said binding level with ISG17 in a control sample from a non-pregnant animal or recently-aborted animal of the same species, or less than that of said binding level of ISG17 in a control sample from a pregnant animal of the same species; said binding level of said antibody with ISG17 in the test sample not greater than that in the control sample from a non-pregnant animal or recently-aborted animal of the same species, or said binding level with ISG17 in the test sample less than that in the control sample from a pregnant animal of the same species being indicative of non-pregnancy; wherein said method does not include a cell-lysing step; (2) if the sheep or cow is determined not to be pregnant, forcing estrus or monitoring for and detecting signs of behavioral estrus; and (3) effecting a second breeding of said sheep or cow. 22. A method detecting non-pregnancy in a sheep or cow by a method comprising: (1) obtaining a sample of blood or serum from said sheep or cow; (2) providing an immunoassay device comprising: (a) at least one absorbent support; (b) an antibody specific to said ISG17 on a said absorbent support; (c) a flow control component on a said absorbent support; (d) means for detecting binding of said antibody with said ISG17; and (e) means for comparing the binding level of ISG17 in the test sample with a background binding level of ISG17 in a control sample from a non-pregnant animal, a pregnant animal or a recently-aborted animal of the same species; (3) flowing the sample into the immunoassay test device into contact with the antibody; (4) detecting a binding level of said antibody with said ISG17 in the test sample not greater than that of said binding level with ISG17 in a control sample from a non-pregnant animal or recently-aborted animal of the same species, or less than that of said binding level of ISG17 in a control sample from a pregnant animal of the same species; said binding level of said antibody with ISG17 in the test sample not greater than that in the control sample from a non-pregnant animal or recently-aborted animal of the same species, or said binding level with ISG17 in the test sample less than that in the control sample from a pregnant animal of the same species being indicative of non-pregnancy; wherein said method does not include a cell-lysing step.