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CN-121995067-A - Dissociating agent and kit for progesterone determination

CN121995067ACN 121995067 ACN121995067 ACN 121995067ACN-121995067-A

Abstract

The invention relates to the field of biotechnology, in particular to a dissociation agent and a kit for progesterone determination. The sample dissociating agent for detecting the progesterone is obtained through optimized screening, comprises hydrocortisone, prednisolone and phthalic acid mono (2-ethylhexyl) ester (MEHP), has high sensitivity to sample detection, good detection linear range and better correlation with a comparison reagent when the components of the dissociating agent are independently acted and combined, has remarkable overall dissociating effect, can release the progesterone from corticosteroid binding protein, albumin and sex hormone binding globulin, is favorable for binding the progesterone with a capture antibody, and can effectively improve sensitivity and accuracy. The invention can be directly added into the reagent component, so that the dissociation process and the immune reaction are carried out simultaneously, the immune reaction is concise and efficient, and the reaction efficiency is improved.

Inventors

  • LI JUN
  • Zhang Kuize

Assignees

  • 北京北方生物技术研究所有限公司

Dates

Publication Date
20260508
Application Date
20260205

Claims (10)

  1. 1. Sample dissociating agent for progesterone detection, comprising hydrocortisone, prednisolone and/or mono (2-ethylhexyl) phthalate.
  2. 2. The sample dissociating agent of claim 1, wherein the concentration of hydrocortisone is 1-10 μg/mL.
  3. 3. The sample dissociating agent of claim 1, wherein the concentration of prednisolone is 10-100 μg/mL.
  4. 4. The sample dissociating agent of claim 1, wherein the concentration of MEHP is 1-10 μg/mL.
  5. 5. The sample dissociation agent of any one of claims 1-4, further comprising a buffer component.
  6. 6. The sample dissociating agent of claim 5, wherein the buffer component comprises a phosphate.
  7. 7. The progesterone detection kit is characterized by comprising the sample dissociating agent and other auxiliary detection reagents according to claims 1-6.
  8. 8. The progesterone assay kit according to claim 7, wherein the additional auxiliary detection reagents comprise progesterone standard, antibodies, immunoisolatory agents, radiolabeled ligands and/or quality control serum.
  9. 9. The progesterone detection kit of claim 8 wherein the antibodies comprise rabbit anti-P antibodies.
  10. 10. The progesterone detection kit of claim 8, wherein the radiolabeled ligand comprises 125 I-P.

Description

Dissociating agent and kit for progesterone determination Technical Field The invention relates to the field of biotechnology, in particular to a dissociation agent and a kit for progesterone determination. Background Progesterone (Progesterone, prog) is a 21-carbon steroid hormone with a molecular weight of 314.5 daltons and is an intermediate metabolite in the synthesis of steroid hormones. However, female progesterone in the blood is mostly produced by the corpus luteum or placenta, while male progesterone levels are very low, mainly produced by the adrenal cortex. Progesterone is an important hormone that plays an important role not only in the regulation of the menstrual cycle but also in maintaining pregnancy. The progesterone can grow glands in uterine mucosa in the later period of menstrual cycle, engorge uterus and thicken endometrium, and is prepared for fertilized egg implantation, which can generate placenta after fertilized egg implantation, reduce excitability of pregnant uterus, inhibit activity, and ensure safe growth of fetus. The progesterone assay is used for determining ovulation, progestogen therapy monitoring and early pregnancy condition evaluation, has a particularly important meaning in judging the luteal function state and aspect, and is an indispensable means for researching the physiology and pathophysiology of the ovary. In the plasma, about 80% -90% of progesterone is combined with corticosteroid-binding globulin, and 10% -15% of progesterone is combined with albumin and sex hormone-binding globulin. Progesterone assay reagents require that progesterone be cleaved off of corticosteroid-binding proteins, albumin, sex hormone binding globulin in order to be detected accurately. The existing progesterone dissociation agent has poor dissociation effect, so that part of low-end blood samples cannot detect effective concentration or have too low measured value, and therefore, the novel progesterone dissociation agent which has good dissociation effect and can be used for accurately detecting progesterone in the low-end blood samples is particularly necessary. Disclosure of Invention In view of the above, the technical problem to be solved by the present invention is to provide a dissociation agent and a kit for progesterone assay. The present invention provides a sample dissociating agent for progesterone detection comprising hydrocortisone, prednisolone and/or mono (2-ethylhexyl) phthalate (MEHP). Specifically, the dissociating agent according to the present invention includes several cases as shown below; case 1, the dissociating agent is hydrocortisone; in the case 2, the dissociating agent is prednisolone; Case 3, wherein the dissociating agent is MEHP; Case 4, the dissociating agent is MEHP and hydrocortisone; Case 5, wherein the dissociating agent is MEHP and prednisolone; Case 6, the dissociating agent is hydrocortisone and prednisolone; Case 7, the dissociating agent is MEHP, prednisolone and hydrocortisone; According to the invention, by comparing and screening the dissociation effects of prednisolone, hydrocortisone, MEHP, di (2-ethylhexyl) phthalate (DEHP for short), 2-bromoestradiol, danazol (daralamin), mesterone and the like on progesterone from a sample, the dissociation effect of hydrocortisone, prednisolone and/or mono (2-ethylhexyl) phthalate (MEHP) is good when acting independently, and the correlation with Beckmann dissociation effect is high and higher than 0.97. Meanwhile, a dissociation combination mode is adopted, after the dissociation agents are combined, the linearity and sensitivity of the calibrator are improved obviously, the correlation between the sample measured value and the comparison reagent is good, and the dissociation effect is improved obviously as a whole. The progesterone can be better released from the corticosteroid binding protein, albumin and sex hormone binding globulin, which is favorable for the combination of the progesterone and the capture antibody, and can effectively improve the sensitivity and the accuracy. Can be directly added into the component serving as a radioimmunoassay reagent to improve the immune response efficiency. In the sample dissociation agent disclosed by the invention, the concentration of hydrocortisone is 1-10 mug/mL, preferably 6 mug/mL-10 mug/mL, and more preferably 2 mug/mL. The concentration of the prednisolone is 10-100 mug/mL, preferably 60 mug/mL-100 mug/mL, and more preferably 20 mug/mL. The concentration of MEHP is 1-10. Mu.g/mL, preferably 8-10. Mu.g/mL, more preferably 4. Mu.g/mL. The sample dissociation agent of the present invention further comprises a buffer component. The buffer component comprises phosphate. The dissociating agent of the present invention further includes a stabilizer, a preservative, and/or other components for maintaining stability of the dissociating agent, which is not limited in the present invention. According to the invention, through screening, other auxiliary material