Systems and methods for detecting animal pregnancy

A method of determining whether a non-human animal is pregnant comprising:

• providing a test kit including at least three test surfaces, each surface being coated with a first biomolecular recognition element specific to a marker of pregnancy, a first reagent solution including a second biomolecular recognition element specific to the marker, a first standard corresponding to a first amount of the marker, and a second standard corresponding to a second amount of the marker that is lower than the first amount,
• obtaining a first sample from the animal after insemination of the animal,
• introducing equal aliquots of the first reagent solution to each of the three surfaces,
• introducing the first standard to a first one of the surfaces,
• introducing the second standard to a second one of the surfaces,
• introducing the sample to a third one of the surfaces,
• allowing the standards and sample to react with the first biomolecular recognition element to bind any of the marker present to the three surfaces, and the second biomolecular recognition element to react with the marker bound to each of the three surfaces, under a substantially constant temperature for a specified time period,
• removing unbound materials, including unbound marker, unbound standards, unbound sample, and unbound first reagent solution, from the three test surfaces,
• introducing to the three surfaces a visual indicator to undergo a visually detectable change, the magnitude of which is related to the amount of the second biomolecular recognition element bound to each surface,
• visually comparing with the naked eye, without the aid of an instrument, under a substantially constant temperature, and after a specified time period, a color intensity of the visual indicator present in the third one of the surfaces with respective color intensities of the visual indicators present in the first and second surfaces,
• determining, based on the visually comparing step, whether a magnitude of the color intensity of the third surface is greater than that of the first surface, is equal to or greater than that of the first surface and equal to or less than that of the second surface, or is less than that of the second surface,
• concluding that the animal is pregnant when the magnitude of the color intensity of the visual indicator of the third surface is greater than that of the first surface,
• concluding that the animal is not pregnant when the magnitude of the color intensity of the visual indicator of the third surface is lower than that of the second surface, and
• concluding that retesting is required of a second sample from the animal, retrieved at a time after the first sample was retrieved, if the magnitude of the color intensity of the visual indicator of the third surface is between the magnitudes of the visual indicators of the first and second surfaces, and
• retesting the second sample to visually determine with the naked eye, without the aid of an instrument, whether the second sample indicates that the cow is pregnant, not pregnant, or another retest is required, wherein the second sample is retrieved from the animal between 24 hours and 7 days after the first sample was retrieved from the animal.