Pregnancy detection

• US 20030224452A1
• Filed: 05/02/2003
• Published: 12/04/2003
• Est. Priority Date: 05/02/2002
• Status: Active Grant

• Current Assignee: Aspenbio Pharma, Inc., University of Wyoming
• Original Assignee: Aspenbio Pharma, Inc.
• Inventors: Landmann, Cathy, Colgin, Mark, Hurst, Roger, Newman, Diane, Roth, J.W.
• Granted Patent: US 7,842,513 B2
• Time in Patent Office: Days
• US Class Current: 435/7.1
• CPC Class Codes: A61D 17/006 for detecting pregnancy of animals
• CPC Class Codes: C07K 16/18 against material from animals or humans
• CPC Class Codes: G01N 2333/4715 Cytokine-induced proteins
• CPC Class Codes: G01N 2800/368 Pregnancy complicated by disease or abnormalities of pregnancy, e.g. preeclampsia, preterm labour
• CPC Class Codes: G01N 33/689 related to pregnancy or the gonads
• CPC Class Codes: Y10S 435/806 Fertility tests
• CPC Class Codes: Y10S 436/807 Apparatus included in process claim, e.g. physical support structures
• CPC Class Codes: Y10S 436/814 Pregnancy
• CPC Class Codes: Y10S 436/906 Fertility tests

Abstract

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, occular 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.

Claims

1. A method for producing a monoclonal antibody comprising

a) providing a polypeptide 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);

b) administering said peptide to a mammal under conditions appropriate for stimulation of an immune response;

c) isolating antibody producing cells from the mammal;

d) fusing the antibody producing cells with immortalizing cells to produce a hybridoma cell line; and

e) screening the resulting hybridoma cell line to identify a cell line secreting antibody having the desired specificity;

said monoclonal antibody capable of binding to said selected polypeptide.

2. A cell of a hybridoma made by the method of claim 1.

3. A monoclonal antibody made by the method of claim 1.

4. The method of claim 1 wherein said polypeptide is produced synthetically.

5. A monoclonal antibody generated with and capable of 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).

6. A polyclonal antibody generated with and capable of 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).

7. A method for producing a polyclonal antibody comprising

a) providing a polypeptide 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);

b) administering said peptide to a mammal under conditions appropriate for the stimulation of an immune response; and

c) isolating the polyclonal antibody from the mammal;

said polyclonal antibody capable of binding to said selected polypeptide.

8. A polyclonal antibody produced by the method of claim 7.

9. The method of claim 7 wherein said polypeptide is produced synthetically.

10. An immunogenic composition comprising 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).

11. The immunogenic composition of claim 10 comprising a polypeptide produced synthetically.

12. An immunoassay test device for detecting the presence of a selected IFN-τ

-induced protein in a sample, said device comprising a first antibody specific to said selected IFN-τ

-induced protein, a support for said first antibody, means for contacting said first antibody with said sample, and a means for detecting binding of said first antibody with said selected IFN-τ

-induced protein.

13. The device of claim 12 wherein said antibody is a monoclonal or polyclonal antibody to a polypeptide 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).

14. The immunoassay test device of claim 12 also comprising means for lysing cells in said sample.

15. The immunoassay test device of claim 12 which is a lateral flow device, also comprising inlet means for flowing a liquid sample into contact with said first antibody.

16. The immunoassay test device of claim 15 also comprising a filter for filtering serum or plasma from a blood sample.

17. The device of claim 12 wherein said indicator comprises a second antibody specific to said selected IFN-τ

-induced protein.

18. The device of claim 17 wherein said second antibody is a polyclonal or monoclonal antibody.

19. The device of claim 17 wherein said second antibody binds a substantially distinct epitope from the first monoclonal antibody.

20. The device of claim 17 wherein said second antibody is selected from the group consisting of 5E9, anti-ISG17-4245, and anti-ISG17-1000.

21. The device of claim 20 wherein said second antibody is capable of binding to said selected IFN-τ

-induced protein while the first antibody is bound to said selected IFN-τ

-induced protein.

22. The device of claim 12 wherein said antibody is selected from the group consisting of:

antibodies made using 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

antibodies made using complete or partial amino acid sequences from mammalian ISG17, Mx, GCP-2, 2′

,5′

oligoadenylate synthetase, β

2-microglobulin, IRF-1, IRF-2, 1-8U, 1-8D, Leu-13/9-27, or COX-2.

23. The device of claim 12 wherein said antibody is selected from the group consisting of 5E9, anti-ISG17-4245, and anti-ISG17-1000.

24. The device of claim 12 wherein the IFN-τ

-induced protein is selected from the group consisting of ISG17, Mx, GCP-2, 2′

,5′

oligoadenylate synthetase, β

2-microglobulin, IRF-1, IRF-2, 1-8U, 1-8D, Leu-13/9-27, and COX-2.

25. The device of claim 12 wherein said IFN-τ

-induced protein is selected from the group consisting of ISG17, Mx, IRF-1, IRF-2, 1-8U, 1-8D, and Leu-13/9-27.

26. The device of claim 12 wherein said IFN-τ

-induced protein is selected from the group consisting of ISG17 and Mx.

27. The device of claim 12 wherein said IFN-τ

-induced protein is ISG17.

28. The device of claim 12 also comprising a housing.

29. The device of claim 12 in the format of a dipstick or a cartridge.

30. A kit comprising the device of claim 12 and instructions for using said device.

32. A method of detecting the presence of a selected IFN-τ

-induced protein comprising;

a) providing the immunoassay test device of claim 12;

b) contacting said antibody with said sample; and

c) reading the results of said means for detecting binding.

33. The method of claim 32 wherein detection of binding indicates pregnancy of a mammal.

34. The method of claim 32 wherein said mammal is an ungulate or non-hoofed ruminant.

35. The method of claim 32 wherein said mammal is selected from the group consisting of cattle, sheep, goats, yaks, water buffaloes, bison, antelopes, gazelles, elk, reindeer, moose, bighorn sheep, giraffes, and camellids including bactrian and dromedary camels, llamas, swine, horses, alpacas, and vicunas.

36. The method of claim 32 performed cowside.

37. The method of claim 32 wherein said IFN-τ

-induced protein is ISG17.

38. The method of claim 32 also comprising comparing said reading of step c) with a reading of normal background levels of said IFN-τ

-induced protein using an equivalent immunoassay test device.

39. The method of claim 32 also comprising collecting said sample from an animal in the field.

31. An immunoassay test device for detecting the presence of a selected IFN-τ

-induced protein in a sample at a level substantially greater than normal background level thereof in said sample, said device comprising an antibody specific to said selected IFN-τ

-induced protein, a support for said antibody, means for contacting said antibody with said sample, an indicator capable of detecting binding of said antibody with said selected IFN-τ

-induced protein; and

means for comparing binding of said antibody in said sample with binding of said antibody in a second sample containing normal background levels of said IFN-τ

-induced protein.

40. A method for making an immunoassay test device comprising:

a) supplying an antibody specific to a selected IFN-τ

-induced protein;

b) attaching said antibody to a support;

c) providing means for contacting said antibody with a sample;

d) providing an indicator for detecting binding of said antibody to said protein; and

e) assembling said support with said antibody attached thereto, said means for contacting said antibody with a sample, and said indicator.

41. The method of claim 40 wherein said antibody is a monoclonal or polyclonal antibody to a polypeptide 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).

42. The method of claim 40 also comprising containing the assembled components of step e) in a housing.

43. The method of claim 40 also comprising assembling the assembled components of step e) into a dipstick or cartridge format.

44. The method of claim 40 also comprising providing means for comparing binding of said antibody in said sample with binding of said antibody in a second sample containing normal background levels of said IFN-τ

-induced protein, and assembling said means for comparing with said antibody, indicator, and support.

45. The method of claim 40 also comprising providing instructions for use of said device and assembling said instructions together with said antibodies, indicator, and support.

46. A method for making an immunoassay test device comprising:

a) supplying a first antibody specific to a selected IFN-τ

-induced protein;

b) attaching, but not immobilizing, said first antibody conjugated to label to a first support;

c) supplying a second antibody specific to said selected IFN-τ

-induced protein;

d) immobilizing said second antibody to a second support;

e) providing means for contacting said first and second antibodies with a sample; and

f) assembling said supports, said means for contacting said antibodies with a sample, and said indicator.

47. The method of claim 46 also comprising

a) attaching and/or immobilizing flow control component on a third support;

and

b) providing means for contacting flow control components with said sample.

48. The method of claim 46 wherein said first monoclonal antibody is 5E9.

49. The method of claim 46 wherein said second antibody is selected from the group consisting of polyclonal antibodies anti-ISG17-4245 and anti-ISG17-1000.

50. The method of claim 46 wherein said label is colloidal gold.

51. A method of detecting pregnancy in an ungulate or other ruminant female animal comprising:

a) obtaining a sample from said animal; and

b) detecting the presence or level of a selected IFN-τ

-induced protein in said sample;

whereby the presence or level of said selected IFN-τ

-induced protein indicates that said animal is pregnant.

52. The method of claim 51 wherein method step b) comprises using an immunoassay test device for detecting the presence or level of a selected IFN-τ

-induced protein in a sample, said device comprising an antibody specific to said selected IFN-τ

-induced protein, a support for said antibody, means for contacting said antibody with said sample, and a means for detecting binding of said antibody with said selected IFN-τ

-induced protein.

53. The method of claim 51 wherein said sample is obtained on a day selected from the group consisting of Day 11, Day 32, and intervening days after breeding.

54. The method of claim 53 wherein said breeding comprises performing artificial insemination.

55. The method of claim 53 wherein said breeding is preceded by forced estrus.

56. The method of claim 51 wherein said sample is obtained on a day selected from Day 15, Day 25, and intervening days after breeding.

57. The method of claim 51 wherein said sample is obtained on a day selected from Day 18, Day 22, and intervening days after breeding.

58. The method of claim 51 wherein said sample is obtained on a day selected from Day 18 or Day 19 after breeding.

59. The method of claim 51 wherein said sample is obtained on Day 18 after breeding.

60. The method of claim 51 wherein said sample is obtained on Day 19 after breeding.

61. The method of claim 51 wherein the method step of detecting the presence of a selected IFN-τ

-induced protein is selected from the group consisting of western blot, sandwich blot, ELISA, dot blot, slot blot, Northern blot, and antibody precipitation.

62. The method of claim 51 wherein the level of said selected IFN-τ

-induced protein indicating the animal is pregnant is substantially above the background level of said selected IFN-τ

-induced protein in a non-pregnant animal.

63. The method of claim 51 wherein said animal is not infected with a virus.

64. The method of claim 51 wherein the IFN-τ

-induced protein is selected from the group consisting of ISG17, Mx, GCP-2, 2′

,5′

oligoadenylate synthetase, β

2-microglobulin, IRF-1, IRF-2, 1-8U, 1-8D, Leu-13/9-27, and COX-2.

65. The method of claim 51 wherein said IFN-τ

-induced protein is selected from the group consisting of ISG17, Mx, IRF-1, IRF-2, 1-8U, 1-8D, and Leu-13/9-27.

66. The method of claim 51 wherein said IFN-τ

-induced protein is selected from the group consisting of ISG17 and Mx.

67. The method of claim 51 wherein said said IFN-τ

-induced protein is not Mx.

68. The method of claim 51 wherein said selected IFN-τ

-induced protein is ISG17.

69. The method of claim 51 wherein said selected IFN-τ

-induced protein is secreted.

70. The method of claim 51 wherein said sample is blood.

71. The method of claim 70 also comprising filtering a blood sample to obtain serum containing said selected IFN-τ

-induced protein.

72. The method of claim 51 wherein said sample is selected from the group comprising cells, whole blood, plasma, serum, milk, saliva, and urine.

73. The method of claim 51 performed cowside.

74. The method of claim 51 wherein said animal is selected from the group consisting of cattle, sheep, goats, yaks, water buffaloes, bison, antelopes, gazelles, elk, reindeer, moose, bighorn sheep, giraffes, and camellids including bactrian and dromedary camels, llamas, swine, horses, alpacas, and vicunas.

75. The method of claim 51 wherein said detecting comprises immunological detecting.

76. The method of claim 75 wherein immunological detecting comprises using an antibody wherein said antibody is selected from the group consisting of:

5E9;

anti-ISG17-4245;

anti-ISG17-1000;

monoclonal and polyclonal antibodies made using 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 made using complete or partial amino acid sequences from mammalian ISG17, Mx, GCP-2, 2′

,5′

oligoadenylate synthetase, β

2-microglobulin, IRF-1, IRF-2, 1-8U, 1-8D, Leu-13/9-27, or COX-2.

77. The method of claim 75 wherein said immunological detecting comprises binding an antibody to said selected IFN-τ

-induced protein and detecting said binding.

78. The method of claim 77 wherein said first antibody is a monoclonal or polyclonal antibody.

79. The method of claim 77 also comprising comparing binding of said antibody in said sample with binding of said antibody in a second sample containing normal background levels of said IFN-τ

-induced protein.

80. The method of claim 79 also comprising binding a second antibody to said selected IFN-τ

-induced protein.

81. The method of claim 77 wherein said antibody is labeled.

82. The method of claim 77 wherein said antibody is selected from the group consisting of 5E9, anti-ISG17-1000, and anti-ISG17-4245.

83. The method of claim 77 also comprising binding a second antibody to said selected IFN-τ

-induced protein.

84. The method of claim 83 wherein said second antibody is selected from the group consisting of anti-ISG17-1000, anti-ISG17-4245, and 5E9.

85. The method of claim 84 wherein said second antibody binds a substantially distinct epitope from the first antibody.

86. A method of detecting non-pregnancy in an ungulate or other ruminant female animal comprising:

a) obtaining a sample from said animal; and

b) detecting the absence of a selected IFN-τ

-induced protein;

whereby the absence of said selected IFN-τ

-induced protein indicates that said animal is not pregnant.

87. A method for determining readiness for breeding comprising

a) performing the method of claim 86;

and

b) forcing estrus or monitoring for and detecting signs of behavioral estrus.

88. A method of determining the pregnancy status of an ungulate or other ruminant female animal, on a day after breeding of said animal, wherein said animal was bred during estrus, said day being a day before the next expected estrus of said animal, said method comprising:

a) obtaining a sample from said animal; and

b) determining pregnancy status of said animal.

89. The method of claim 88 wherein step b) comprises assaying for the presence or absence of a selected IFN-τ

-induced protein.

90. A method for determining readiness for breeding of a female animal of a species utilizing IFN-τ

mediated signaling for maternal recognition of pregnancy comprising;

a) detecting the absence of a selected IFN-τ

-induced protein in a sample taken from said animal on a day of said animal'"'"'s estrus cycle during which the presence of said selected IFN-τ

-induced protein is detectable if said animal is pregnant; and

b) determining said animal is in estrus;

thereby determining readiness for breeding.

91. The method of claim 90 wherein method step b) comprises forcing estrus or monitoring for and detecting behavioral estrus.

92. The method of claim 91 wherein forcing estrus in said animal comprises a method step selected from the group consisting of administering one prostaglandin injection and administering a regimen of two GnRH injections with one prostaglandin injection.

93. The method of claim 91 also comprising monitoring for and detecting signs of behavioral estrus wherein said determining comprises forcing said animal into heat.

94. The method of claim 90 wherein method step a) comprises using an immunoassay test device for detecting the presence of a selected IFN-τ

-induced protein in a sample, said device comprising an antibody specific to said selected IFN-τ

-induced protein, a support for said antibody, means for contacting said antibody with said sample, and a means for detecting binding of said antibody with said selected IFN-τ

-induced protein.

95. The method of claim 90 wherein said breeding comprises artificial insemination.

96. The method of claim 90 wherein said breeding comprises natural insemination.

97. The method of claim 90 wherein said animal is bovine.

98. The method of claim 90 wherein said day is selected from the group consisting of Day 11, Day 32, and intervening days.

99. The method of claim 90 wherein said day is selected from the group consisting of Day 15, Day 25, and intervening days.

100. The method of claim 90 wherein said day is selected from the group consisting of Day 18, Day 22, and intervening days.

101. The method of claim 90 wherein said day is selected from the group consisting of Day 18, Day 19.

102. The method of claim 90 wherein said day is Day 18.

103. The method of claim 90 wherein said day is Day 19.

104. The method of claim 90 wherein the selected IFN-τ

-induced protein is selected from the group consisting of ISG17, Mx, GCP-2, 2′

,5′

-oligoadenylate synthetase, β

2-microglobulin, IRF-1, IRF-2, 1-8U, 1-8D, Leu-13/9-27, and COX-2.

105. The method of claim 90 wherein the selected IFN-τ

-induced protein is ISG17.

106. The method of claim 90 wherein the sample is selected from the group consisting of cells, whole blood, plasma, serum, milk, saliva, and urine.

107. The method of claim 90 wherein the method step of detecting the absence of a selected IFN-τ

-induced protein in a sample from said animal comprises

a) obtaining a sample from said animal; and

b) detecting the absence of a selected IFN-τ

-induced protein;

whereby the absence of said selected IFN-τ

-induced protein indicates that said animal is not pregnant.

108. The method of claim 107 wherein said detecting comprises immunological detecting.

109. The method of claim 108 wherein said immunological detecting comprises contacting said sample with a first polyclonal or monoclonal antibody to said selected IFN-τ

-induced protein and detecting failed binding.

110. The method of claim 109 wherein said first monoclonal or polyclonal antibody is labeled.

111. The method of claim 109 also comprising binding a second antibody to said selected IFN-τ

-induced protein.

116. The method of claim 90 wherein said animal is selected from the group consisting of cattle, sheep, goats, yaks, water buffaloes, bison, antelopes, gazelles, elk, reindeer, moose, bighorn sheep, giraffes, and camellids including bactrian and dromedary camels, llamas, swine, horses, alpacas, and vicunas.

117. The method of claim 90 performed on plurality of female animals.

112. A method for determining readiness for breeding of a female animal of a species utilizing IFN-τ

-mediated signaling for maternal recognition of pregnancy comprising;

a) obtaining a sample from said animal; and

b) detecting the absence of an elevated level above background levels of said selected IFN-τ

-induced protein;

whereby the absence of said elevated levels of said selected IFN-τ

-induced protein indicates that said animal is not pregnant.

113. The method of claim 112 wherein said detecting comprises immunological detecting.

114. The method of claim 113 wherein immunological detecting comprises contacting said sample with a monoclonal or polyclonal antibody to said selected IFN-τ

-induced protein and detecting binding substantially the same as binding of said monoclonal or polyclonal antibody to a sample from a non-pregnant animal.

115. The method of claim 114 wherein said monoclonal or polyclonal antibody is selected from the group consisting of:

anti-ISG17-4245;

anti-ISG17-1000;

5E9;

monoclonal and polyclonal antibodies made using 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

antibodies made using complete or partial amino acid sequences from mammalian ISG17, Mx, GCP-2, 2′

,5′

-oligoadenylate synthetase, β

2-microglobulin, IRF-1, IRF-2, 1-8U, 1-8D, Leu-13/9-27, or COX-2.

118. A method for breeding a plurality of female animals of a species utilizing IFN-τ

-mediated signaling for maternal recognition of pregnancy comprising;

a) optionally estrus presynchronizing said animals;

b) estrus synchronizing said animals;

c) breeding said animals;

d) pregnancy testing said animals to determine non-pregnant animals;

e) estrus resynchronizing said non-pregnant animals; and

f) rebreeding said non-pregnant animals.

119. The method of claim 118 wherein method step d) comprises using an immunoassay test device for detecting the presence of a selected IFN-τ

-induced protein in a sample, said device comprising an antibody specific to said selected IFN-τ

-induced protein, a support for said antibody, means for contacting first antibody with said sample, and a means for detecting binding of said antibody with said selected IFN-τ

-induced protein.

120. The method of claim 118 wherein said plurality of animals is bovine.

121. The method of claim 118 wherein said estrus synchronizing is ovulation synchronizing.

122. The method of claim 118 wherein the method step d) of pregnancy testing said animals comprises:

a) obtaining a sample from each said animals on a day of said animal'"'"'s estrus cycle during which the presence of a selected IFN-τ

-induced protein would indicate said animal is pregnant; and

b) detecting the absence of said selected IFN-τ

-induced protein in each said sample;

whereby detecting the absence of said selected IFN-τ

-induced protein in said sample indicates that said animal is non-pregnant.

123. The method of claim 118 wherein said pregnancy testing occurs on Day 18.

124. The method of claim 118 wherein said estrus resynchronizing comprises injecting prostaglandin.

125. The method of claim 124 wherein said injecting prostaglandin occurs on Day 18.

126. A method of breeding a female animal of a species utilizing IFN-τ

-mediated signaling for maternal recognition of pregnancy, said animal having been previously bred during estrus, comprising;

a) pregnancy testing said animal to determine non-pregnancy;

b) determining said non-pregnant animal is in estrus; and

c) breeding said non-pregnant animal.

127. The method of claim 126 wherein method step c) comprises performing artificial insemination

128. The method of claim 126 wherein method step b) comprises forcing estrus or monitoring for and detecting behavioral estrus.

129. The method of claim 128 wherein said forcing step comprises injecting prostaglandin.

130. The method of claim 129 comprising injecting about 25 mg of prostaglandin.

131. The method of claim 126 wherein method step c) occurs within the same estrus cycle as the previous breeding.

132. The method of claim 126 wherein method step a) comprises:

1) obtaining a sample from said animal on a day of said animal'"'"'s estrus cycle during which the presence of a selected IFN-τ

-induced protein would indicate if said animal is pregnant; and

2) detecting the absence of said selected IFN-τ

-induced protein in said sample;

whereby detecting the absence of said selected IFN-τ

-induced protein in said sample indicates that said animal is non-pregnant.

133. The method of claim 132 wherein said sample is taken on a day selected from the group consisting of Day 11, Day 32, and intervening days.

134. The method of claim 132 wherein said sample is taken on a day selected from the group consisting of Day 15, Day 22, and intervening days.

135. The method of claim 132 wherein said sample is taken on a day selected from the group consisting of Day 18 and Day 19

136. The method of claim 132 wherein said forcing estrus comprises:

a) injecting GnRH;

b) waiting about 7 days;

c) injecting prostaglandin; and

d) waiting until observation of behavioral estrus or the morning of the third day after performing method step c).

137. A method of rebreeding an ungulate or other ruminant female animal, wherein said animal was bred during a first estrus, comprising:

a) determining non-pregnant status of said animal;

b) forcing estrus; and

c) rebreeding said animal;

wherein said rebreeding occurs before the time when an unforced second estrus is likely to occur.