Rapid test kit for the early detection of pregnancy in pigs using fecal examination

Abstract

A rapid test kit for the early detection of pregnancy in pigs using fecal examination, including: an extraction container, wherein a fecal sample, an extraction unit and a progesterone-containing extraction unit are contained in the extraction container; and an examination unit located adjacent to the extraction container, wherein the progesterone-containing extraction unit is placed on the examination unit to determine whether the sow is in a state of early pregnancy.

Description

Field of invention

[0001] The present invention relates to a rapid test kit and in particular a rapid test kit for the early detection of pregnancy in pigs by means of fecal examination. State of the art

[0002] A sow's production cycle comprises the phases of mating, pregnancy, farrowing, lactation, estrus, and rebreeding. If an error occurs at any of these stages, the sow's piglet production rate will inevitably be affected, resulting in economic losses for the farmer. The average gestation period for a sow is 114 days, although approximately 10% to 30% of sows do not become pregnant after mating. If non-pregnant sows are not detected early, they will consume feed for an average of another 60 days before this is discovered.

[0003] It is difficult to determine externally whether a sow is pregnant or not. Often, a false pregnancy is only discovered shortly before birth. This situation is not uncommon in pig farming operations and leads not only to wasted feed during this period and increased housing costs, but also to a reduction in the annual number of piglets per sow and thus to a decrease in agricultural yields. Therefore, the reliable diagnosis of sow pregnancy is a pressing problem for the relevant industries.

[0004] According to current technology, various methods exist for the early detection of pregnancy in pigs. Ultrasound examinations and hormonal blood tests are among the most common. However, ultrasound examinations require the purchase of expensive ultrasound equipment and the training of operators, which is associated with higher costs. Hormonal blood tests primarily determine pregnancy based on the progesterone level (P4). Due to individual differences between the pigs examined and possible reproductive problems, a false pregnancy (pseudopregnancy), for example, caused by a persistent corpus luteum producing progesterone, may not be detected. Therefore, the accuracy is not 100%.

[0005] Furthermore, all conventional methods for the early detection of pregnancy in pigs require a waiting period of more than 20 days after mating before a diagnosis is possible, regardless of whether ultrasound measurements, hormonal blood tests, or other methods are used. If, after these 20 days, it is determined that there is no early pregnancy, feed is unnecessarily consumed by the sow during this period, leading to increased husbandry costs.

[0006] How the aforementioned disadvantages can be avoided and problems solved is therefore of great importance to the inventor and a technical problem that needs to be solved. Object of the invention

[0007] In order to effectively solve the above problems, the main objective of the present invention is to provide a rapid test kit for the early detection of pregnancy in pigs by means of fecal examination, which can significantly reduce husbandry costs.

[0008] A secondary objective of the present invention is to provide a rapid test kit for the early detection of pregnancy in pigs by means of fecal examination, with which the time until the determination of early pregnancy can be significantly reduced.

[0009] A secondary objective of the present invention is to provide a rapid test kit for the early detection of pregnancy in pigs by means of fecal examination, with which agricultural yields can be significantly increased.

[0010] A secondary objective of the present invention is to provide a rapid test kit for the early detection of pregnancy in pigs by means of fecal examination, with which the accuracy of early pregnancy detection can be significantly improved.

[0011] To solve the above-mentioned problems, the present invention provides a rapid test kit for the early detection of pregnancy in pigs by means of fecal examination, comprising an extraction container and an examination unit, wherein a fecal sample, an extraction unit and a progesterone-containing extraction unit are received in the extraction container, the examination unit is arranged adjacent to the extraction container and the progesterone-containing extraction unit is arranged on the examination unit to determine whether the sow is in the state of early pregnancy.

[0012] In one embodiment, the extraction unit is a mixed solution of phosphate-buffered salt solution and water.

[0013] In one embodiment, the pH value of the extraction unit is in the range of 6.8 to 7.6.

[0014] In one embodiment, one of the immunochromatographic rapid test strips is selected as the unit of investigation, wherein the immunochromatographic rapid test strip is a rapid test strip with colloidal gold.

[0015] In one embodiment, the testing unit is used to determine whether the progesterone concentration in the fecal sample is higher than 30 ng / ml, in order to use this as a diagnostic basis for the early pregnancy of the sow.

[0016] In one embodiment, the method for measuring progesterone concentration is carried out using immunochromatography.

[0017] The structural design of the present invention provides for the use of a fecal sample obtained from the sow between 10 and 18 days after mating. After mixing the fecal sample with the extraction unit, the mixture undergoes steps such as shaking, centrifugation, and standing to release a progesterone-containing extraction unit from the solution. Subsequently, impurities are removed by means of a filter membrane to obtain a sample solution for rapid immunoassay analysis. This sample solution for rapid immunoassay analysis is introduced into an immunochromatographic rapid test instrument and driven by capillary action, so that the sample solution flows along the assay unit, reacts with the colloidal gold-labeled antibody layer, and then passes through the test line and control line areas.The color intensity is evaluated visually or using an optical reader, whereby the relative progesterone concentration in the sample is assessed based on the color intensity to determine whether the sow is in early pregnancy. In this way, the disadvantages of conventional testing methods can be overcome, where the detection of early pregnancy takes too long and thus incurs additional feed costs during the rearing of the sow. Furthermore, the rapid test kit according to the invention eliminates the disadvantage of conventional testing methods of not being able to reliably rule out a false pregnancy, achieving an accuracy of almost 100%. Brief description of the drawings Fig. Figure 1 shows a schematic view of the structure according to the present invention; Fig.2 shows a flowchart of the steps according to the present invention; Fig. Figure 3 shows a diagram of the change in progesterone concentration in the feces of sows during a period of 21 days after mating according to the present invention. Detailed description of the exemplary implementations

[0018] To better understand the above tasks, features of the structure and functions of the present invention, preferred embodiments are described in more detail below with reference to the accompanying drawings.

[0019] The following section presents various applicable embodiments of the rapid test kit according to the invention for the early detection of pregnancy in pigs by means of fecal examination, and describes them in detail with reference to the accompanying drawings. It is understood that the present invention is not limited to the embodiment presented, the drawings, or the detailed descriptions.

[0020] Furthermore, it should be clear to those skilled in the art that the foregoing description merely explains the embodiments mentioned and the accompanying drawings and is not intended to limit the scope of the claims. A person skilled in the art can make numerous modifications to the disclosed embodiments without departing from the spirit or scope of this invention. All equivalent modifications and changes that can be made in accordance with the technical concepts of the present invention fall within the scope of protection of the present invention.

[0021] Furthermore, the orienting terms mentioned in the following embodiments, such as "top, bottom, left, right, front, and back," serve only to refer to the directions in the accompanying drawings. Therefore, these orienting terms should not be interpreted as limiting the scope of protection of the present invention. It should be noted that similar reference numerals and letters denote similar elements in the following figures.

[0022] It will be on Fig.Reference is made to Figure 1, which shows a schematic view of the setup of the rapid test kit according to the invention for the early detection of pregnancy in pigs by means of fecal examination. As shown, a rapid test kit for the early detection of pregnancy in pigs by means of fecal examination comprises an extraction container 1 and an examination unit 2, wherein a fecal sample 10, an extraction unit 11 and a progesterone-containing extraction unit 12 are contained in the extraction container 1. It should be noted that the extraction unit 11 is a mixed solution of phosphate-buffered saline (PBS) and water, wherein the pH value of the extraction unit 11 is in the range of 6.8 to 7.6.

[0023] Test unit 2 is located adjacent to extraction container 1, and the progesterone-containing extraction unit 12 is positioned on top of test unit 2. Test unit 2 uses one of the immunochromatographic rapid test strips, specifically a rapid test strip containing colloidal gold. Test unit 2 is used to determine whether the progesterone concentration in fecal sample 10 is higher than 30 ng / ml, in order to use this as a diagnostic basis for early pregnancy in sows. The progesterone concentration is measured using immunochromatography.

[0024] It will be on Fig. Reference is made to Figure 2, which shows a flowchart of the steps according to the present invention. As shown, a method for the early detection of pregnancy in pigs by means of fecal examination comprises the following steps:

[0025] Step S1: Using a sampling device, a fecal sample is taken from a sow that has already mated, within a period of 10 to 18 days after mating;

[0026] First, feces are collected using a sampling device from a sow in which mating has already taken place, within a period of 10 to 18 days after mating, and are used as fecal sample 10 for the present investigation.

[0027] Step S2: The stool sample is placed in an extraction container, and an extraction unit is added and mixed to create a solution. The solution is then subjected to the steps of shaking, centrifuging, and standing to release the solid-phase steroid hormones contained in the stool sample into the solution to form a progesterone-containing extraction unit.

[0028] The collected fecal sample 10 is then directly placed into the extraction vessel 1, and the extraction unit 11 is added to the extraction vessel 1, so that the fecal sample 10 and the extraction unit 11 are mixed to form a solution. It should be noted that the extraction unit 11 is a mixture of phosphate-buffered saline and water, with a pH value of the extraction unit 11 in the range of 6.8 to 7.6. After the successive steps of shaking, centrifuging, and standing, the solid-phase steroid hormones contained in the fecal sample 10 are released into the solution, forming a progesterone-containing extraction unit 12.

[0029] Step S3: The progesterone-containing extraction unit is filtered using a filter membrane to remove impurities. This produces a sample solution for rapid analysis by immunoassay;

[0030] Subsequently, a filter membrane with a pore size of no more than 0.22 µm is used to filter impurities from the progesterone-containing extraction unit 11, so that after the filtration step it serves as a sample solution for rapid analysis by immunoassay.

[0031] Step S4: The sample solution for rapid immunoassay analysis is introduced into an immunochromatographic rapid test instrument, the instrument comprising an antibody layer labeled with colloidal gold, at least one test line (T) and one control line (C), wherein a progesterone-corresponding reactant is immobilized on the test line; The sample solution for rapid immunoassay analysis is introduced into the immunochromatographic rapid test instrument to perform a sample analysis. The immunochromatographic rapid test instrument comprises an antibody layer labeled with colloidal gold, a test line (T) and a control line (C), wherein a progesterone-corresponding reactant is immobilized on the test line (T).

[0032] Step S5: Driven by capillary action, the sample solution for rapid analysis using immunoassay flows along a testing unit, reacts with the antibody layer labeled with colloidal gold and then passes through the areas of the test line and the control line, whereby the color intensity is evaluated visually or by means of an optical reader;

[0033] Driven by capillary action, the sample solution for rapid analysis by immunoassay gradually spreads along the test unit 2, reacts with the antibody layer labeled with colloidal gold, and then passes through the test line (T) and control line (C) areas. The displayed color intensity is then visually assessed. Alternatively, the color intensity can also be determined using an optical reader. In the present embodiment, one of the immunochromatographic rapid test strips is selected as test unit 2, wherein the immunochromatographic rapid test strip is a rapid test strip containing colloidal gold.

[0034] Step S6: Based on the displayed reaction color or its optical density intensity, the relative progesterone concentration in the fecal sample is assessed, and on the basis of the progesterone concentration, it is determined whether the sow is in a state of early pregnancy;

[0035] Finally, the relative progesterone concentration in fecal sample 10 is assessed by observing and analyzing the reaction color displayed on the testing unit 2 or its optical density intensity for rapid immunoassay analysis. The progesterone concentration is used to determine whether the sow is in early pregnancy. If the measured progesterone concentration is above 30 ng / ml, the sow is diagnosed as early pregnant. Conversely, if the measured progesterone concentration is below 30 ng / ml, the sow is diagnosed as not pregnant.

[0036] Thus, the inventive method uses a fecal sample obtained from the sow between 10 and 18 days after mating. After mixing the fecal sample 10 with the extraction unit 11, the mixture undergoes steps such as shaking, centrifugation, and standing, resulting in the release of a progesterone-containing extraction unit 12 from the solution. Subsequently, impurities are removed by means of a filter membrane to obtain a sample solution for rapid immunoassay analysis. This sample solution for rapid immunoassay analysis is introduced into an immunochromatographic rapid test instrument and driven by capillary action, causing the sample solution to flow along the testing unit 2, react with the antibody layer labeled with colloidal gold, and then pass through the test line and control line areas.The color intensity is evaluated visually or using an optical reader, whereby the relative progesterone concentration in the sample is assessed based on the color intensity to determine whether the sow is in early pregnancy. The examination method according to the invention allows an examination to be completed within 24 hours of taking the fecal sample from the sow, thus enabling an immediate diagnosis of early pregnancy. Furthermore, it is possible to... Fig. 3 shows that the progesterone concentration determined by the inventive examination method in samples obtained from fecal samples taken on the 10th day after mating reaches a relative peak value on average.

[0037] In this way, the disadvantages of conventional examination methods can be overcome, where determining early pregnancy takes too long and thus incurs additional feed costs during the rearing of the sow. Furthermore, it can be seen from Table 1 below that the examination method according to the invention eliminates the disadvantage of conventional examination methods of not being able to reliably rule out a false pregnancy, achieving an accuracy of almost 100%. Table 1 sample Sow feces after mating Days after mating 10 11 12 13 14 15 16 17 18 pregnant / number 12 5 10 9 25 14 8 8 3 not pregnant / number 0 0 0 0 1 0 2 1 0 total / number 12 5 10 9 26 14 10 9 3 accuracy 98%

[0038] In summary, the present invention has the following advantages over the prior art: 1. Significant reduction in keeping costs; 2. Significant reduction in the examination time for determining early pregnancy in pigs; 3. Significant increase in the earnings of livestock farmers; 4. Significant improvement in the accuracy of determining early pregnancy. Reference symbol list 1 extraction container 10 stool sample 11 extraction units 12 progesterone-containing extraction units 2 examination units S1-S6 steps

Claims

[1] A rapid test kit for the early detection of pregnancy in pigs by means of fecal examination, comprising: an extraction container, wherein a fecal sample, an extraction unit and a progesterone-containing extraction unit are contained in the extraction container; and an examination unit located adjacent to the extraction container, wherein the progesterone-containing extraction unit is placed on the examination unit to determine whether the sow is in a state of early pregnancy. [2] Rapid test kit according to claim 1, wherein the extraction unit is a mixed solution of phosphate-buffered saline and water. [3] Rapid test kit according to claim 2, wherein the pH value of the extraction unit is in the range of 6.8 to 7.

6. [4] Rapid test kit according to claim 1, wherein one of the immunochromatographic rapid test strips is selected as the unit of analysis, the immunochromatographic rapid test strip being a rapid test strip containing colloidal gold. [5] Rapid test kit according to claim 4, wherein the testing unit determines whether the progesterone concentration in the fecal sample is higher than 30 ng / ml, in order to use this as a diagnostic basis for the early pregnancy of the sow. [6] Rapid test kit according to claim 5, wherein the method for measuring the progesterone concentration is carried out by immunochromatography.

Images