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CN-121978087-A - Early cancer detection reagent, kit and preparation method and application thereof

CN121978087ACN 121978087 ACN121978087 ACN 121978087ACN-121978087-A

Abstract

The invention relates to the technical field of biomedicine, in particular to a reagent and a kit for early detection of cancer, and a preparation method and application thereof. The early cancer detection reagent provided by the invention is designed by two steps of a pretreatment reagent and a FeCl 3 chromogenic reagent, the pH value of the pretreatment reagent can be adjusted to be 5-6 in a proper range of the reaction of Fe 3+ and the p-hydroxybenzene-R metabolite, ammonia (NH 3 +H + →NH 4 + and Fe 3+ are prevented from reacting) is fixed in an acidic environment, protein in urine is precipitated, the core interference of the ammonia and the protein in the urine is effectively eliminated, meanwhile, the FeCl 3 is utilized to realize the specific detection of the p-hydroxybenzene-R metabolite, the operation is simple, and the report result is rapid, so that the early cancer detection reagent is suitable for laboratories, physical examination screening and home self-detection. The preparation method of the early cancer detection reagent provided by the invention is simple, low in cost, good in application effect and high in accuracy.

Inventors

  • LI RUNYU
  • JI XIAOLING

Assignees

  • 宝康仁达生物技术(北京)有限公司

Dates

Publication Date
20260505
Application Date
20251217

Claims (10)

  1. 1. The early cancer detection reagent is characterized by comprising a pretreatment reagent and a FeCl 3 chromogenic reagent, wherein the pretreatment reagent comprises trichloroacetic acid and citric acid.
  2. 2. The reagent for early detection of cancer according to claim 1, wherein the concentration of trichloroacetic acid in the pretreatment reagent is 7 to 10W/v%, the molar concentration of citric acid in the pretreatment reagent is 0.07 to 0.1mol/L, and the concentration of FeCl 3 in the FeCl 3 coloring reagent is 2 to 5W/v%, and the pH is 1 to 2.
  3. 3. The reagent for early detection of cancer according to claim 1, wherein the subject comprises a p-hydroxyphenyl-R metabolite in urine, preferably wherein the p-hydroxyphenyl-R metabolite comprises p-hydroxyphenylpyruvic acid.
  4. 4. A method for preparing the reagent for early detection of cancer according to any one of claims 1 to 3, comprising the steps of: The preparation of the pretreatment reagent comprises weighing trichloroacetic acid and citric acid, and dissolving in water solution to obtain the pretreatment reagent; The preparation method of the FeCl 3 chromogenic reagent comprises the steps of adding dilute hydrochloric acid and EDTA disodium solution into FeCl 3 aqueous solution to obtain FeCl 3 chromogenic reagent, wherein the pH value of the FeCl 3 aqueous solution is 1-2 after the addition of the dilute hydrochloric acid; The pretreatment reagent and the FeCl 3 chromogenic reagent are packaged independently.
  5. 5. The method according to claim 4, wherein the disodium EDTA solution is added in an amount of 0.05mL-0.1mL per 100mL of FeCl 3 chromogenic reagent, and the disodium EDTA solution is added in a concentration of 0.01W to 0.05W/V%.
  6. 6. A kit for early detection of cancer is characterized in that the effective components comprise early detection reagents of cancer, wherein the early detection reagents of cancer are prepared by the early detection reagents of cancer according to any one of claims 1-3 or the preparation method of claim 4 or 5.
  7. 7. The early cancer detection reagent according to any one of claims 1 to 3 or the early cancer detection reagent prepared by the preparation method according to claim 4 or 5 or the early cancer detection kit according to claim 6 for use in cancer screening, wherein optionally the sample to be tested comprises urine.
  8. 8. A method for using a reagent for early detection of cancer is characterized in that the reagent for early detection of cancer is the reagent for early detection of cancer prepared by any one of claims 1 to 3 or the reagent for early detection of cancer prepared by the preparation method of claim 4 or 5 or the kit for early detection of cancer of claim 6, and comprises the steps of first mixing fresh middle urine with the pretreatment reagent, first standing and filtering to obtain filtrate, adding the FeCl 3 chromogenic reagent into the filtrate for second mixing and second standing, and observing the color change of the solution; If the solution is bluish violet, light violet or light green, judging that the urine contains p-hydroxybenzene-R metabolites, and the cancer cells grow in the subject; if the color of the filtrate after the pretreatment is light yellow or colorless and no bluish violet or green color appears, the urine is judged to contain no p-hydroxybenzene-R metabolite, and no cancer cells grow in the subject.
  9. 9. The method of claim 8, wherein the first mixing time is 1-5min, the first standing time is 10-15min, the second mixing time is 0.5-1min, and the second standing time is 0.5-1min.
  10. 10. The method according to claim 8, wherein the volume ratio of the fresh mid-stream urine to the pretreatment reagent is 3-6:1-3, and 0.15mL-0.25mL of FeCl 3 chromogenic reagent is added to each milliliter of filtrate.

Description

Early cancer detection reagent, kit and preparation method and application thereof Technical Field The invention relates to the technical field of biomedicine, in particular to a reagent and a kit for early detection of cancer, and a preparation method and application thereof. Background Cancer, a serious disease that seriously threatens human life and health in the global area, has a therapeutic effect closely related to the discovery timing. However, in the current clinical common tumor detection method, an obvious short plate exists in the key link of early discovery, and the requirements of cancer prevention and radical treatment are difficult to meet, so that the research and development of early cancer detection reagents are very important. The existing clinical detection method of cancer has the dilemma of 'early discovery', and the existing clinical widely applied tumor detection methods, such as imaging examination (CT, MRI, ultrasound and the like) and conventional tumor marker detection, have obvious limitations in early detection of cancer, and are difficult to realize effective identification in preclinical stages. Such as imaging, is not early-stage when "seen". Imaging requires that the tumor develop to a volume (typically a few millimeters in diameter or more) to be clearly identified. When tumors are found by means of CT, ultrasound, etc., cancer cells often have undergone longer proliferation in the body, may have entered middle and late stages, and even have undergone local infiltration or distant metastasis, missing the best opportunity for intervention. For example, early stage lesions of lung cancer are smaller, and conventional chest CT has a lower detection rate for tiny lesions less than 1 cm in diameter, and at this time cancer cells may already be somewhat invasive. And the specificity and the sensitivity are insufficient as in the conventional tumor marker detection. The detection of common tumor markers (such as carcinoembryonic antigen CEA, alpha fetoprotein AFP and the like) has the problem of low specificity and sensitivity. On the one hand, part of benign diseases may lead to elevated tumor markers, with "false positive" results, causing unnecessary panic, and on the other hand, many early cancer patients have tumor marker levels that are not significantly abnormal, resulting in "false negative" and missed diagnosis. For example, in early gastric cancer patients, only about 20% -30% of people have raised carcinoembryonic antigen, which is difficult to be a reliable basis for early diagnosis. As well as tissue biopsies, biopsies are traumatic and have the limitation of difficult sampling. Tissue biopsy is a gold standard for tumor diagnosis, but the method belongs to invasive operation, may bring pain, bleeding and other risks to patients, has a limited sampling range, is difficult to accurately acquire pathological tissues for tumors with hidden positions or tiny volumes, and cannot realize the purpose of early screening. Thus, the shift of cancer from "treatment" to "prevention" is critical. The core pain point of cancer treatment is "late found", while early detection reagents can break through the bottleneck of the prior art, realize accurate identification in the preclinical stage of cancer (i.e. the stage in which tumor cells have abnormal proliferation but have not formed obvious lesions or caused clinical symptoms), and provide possibility for "preventive" intervention of cancer, the necessity of which is mainly manifested in the following aspects. Firstly, realize "preclinical discovery", break "late dilemma". The development and progression of cancer is a lengthy process, often taking years or even tens of years from the mutation of cancer cells to the formation of clinically detectable tumors. The early detection reagent can discover abnormality in the preclinical stage that the tumor does not form obvious solid focus and has no clinical symptoms by capturing trace tumor related biomarkers (such as circulating tumor DNA, microRNA, tumor related metabolites and the like) in blood and body fluid, greatly advances a detection window and radically changes the passive situation of 'late discovery'. Secondly, the treatment difficulty is reduced, and the cure possibility is improved. Clinical studies show that the treatment difficulty of early cancers is far lower than that of middle and late cancers, and the cure probability is extremely high. For example, the 5-year survival rate of early cervical cancer can reach more than 90%, but the 5-year survival rate of late cervical cancer is less than 30%, and after standard treatment, the 5-year survival rate of early breast cancer can reach about 95%, but the 5-year survival rate of late breast cancer is only 20% -40%. The early detection reagent helps patients find lesions in the stage of cancer germination, at the moment, tumor cells are not spread yet, reversion or radical treatment can be realized by simple means such as