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Kit for early diagnosis of bladder cancer and preparation method of kit

A technology for bladder cancer and analytical reagents, applied in measuring devices, instruments, scientific instruments, etc., can solve problems such as false positives, and achieve the effects of simple operation, high sensitivity, and rapid detection

Active Publication Date: 2013-04-03
INSITUTE OF BIOPHYSICS CHINESE ACADEMY OF SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The sensitivity reaches 57-83%, and the specificity reaches 60-92%. The specificity of patients with hematuria can only reach 46%. In addition, benign prostatic hyperplasia, kidney stones and urinary tract infection all affect the specificity of BTA detection; Immunocyt kit passed Immunofluorescence detection of urine exfoliated cells, although the sensitivity and specificity are very high, in order to achieve high accuracy, a large number of urine exfoliated cells are required, so it has high application value in the treatment and prognosis of bladder cancer patients. However, the application in early diagnosis is unsatisfactory; among them, NMP22 is currently the most widely used bladder cancer marker, and the marketization of ELISA kits has realized the convenience and rapid detection of early diagnosis of bladder cancer, and the detection sensitivity and specificity are also high. In the case of viral infection, kidney / bladder stones, etc., false positives may still occur; the UroVysion kit uses a multi-target fluorescence in situ hybridization (FISH) technology, although the sensitivity and specificity have been greatly improved, and the US FDA has approved it. However, similar to the Immunocyt kit, a large number of urine exfoliated cells are required

Method used

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  • Kit for early diagnosis of bladder cancer and preparation method of kit
  • Kit for early diagnosis of bladder cancer and preparation method of kit
  • Kit for early diagnosis of bladder cancer and preparation method of kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1 Preparation of the chemiluminescent immunoassay kit for the human urinary bladder cancer tumor marker AG-α3β1 of the present invention

[0046] 1. Preparation of standard products

[0047] This method uses horseradish peroxidase to catalyze the luminol and hydrogen peroxide chemiluminescence system, and uses a chemiluminescence instrument to detect the amount definition of AG-α3β1 contained in the sample with a luminescence value of 6685 (that is, the Cut off value selected by this method) 1U / mL. This luminescence value is also equal to the chemiluminescence value of a human bladder cancer cell line (EJ cell) at a concentration of 1000 cells / mL.

[0048] Collect a large number of urine samples from bladder cancer patients with high chemiluminescence values ​​and normal human urine samples (the normal human urine does not contain AG-α3β1), and use normal human urine samples to gradually dilute the bladder cancer patient urine samples, The concentration gradie...

Embodiment 2

[0086] Example 2 Method of using the bladder cancer tumor marker AG-α3β1 chemical method photoimmunoassay assay kit of the present invention

[0087] 1. Sample pretreatment

[0088] Human urine is collected and analyzed directly without any other special treatment.

[0089] 2. Detection steps

[0090] Before using this kit for detection, it is necessary to take out the standard prepared in Example 1, the coated microwell plate, the enzyme-labeled monoclonal antibody solution and each buffer solution, let it stand at room temperature, and then use it after equilibrating to room temperature ; Adjust the incubator or water bath to 37°C; prepare a suitable micro-sampler and corresponding tips and check whether the chemiluminescence instrument is working normally.

[0091] The specific operation steps for using this kit are as follows:

[0092] 1) Take out the components in the kit and equilibrate to room temperature;

[0093] 2) Place the slats required for the experiment on t...

Embodiment 3

[0102] The methodological index of embodiment 3 kits of the present invention

[0103] The test kit prepared in Example 1 is tested according to the conventional manufacturing and testing procedures in the art, and the results are as follows:

[0104] 1. The standard curve of this kit

[0105] This kit uses horseradish peroxidase to catalyze the luminol and hydrogen peroxide chemiluminescence system, and uses a chemiluminescence instrument to detect the luminescence value and draw a standard curve, see the appendix figure 1 , where, I is the luminous intensity, C is the concentration of the standard cell lysate (in U / mL), and the correlation coefficient R 2 =1, Y=1240.5+5439.54X.

[0106] 2. Kit sensitivity experiment

[0107] Sensitivity is the concentration that can be distinguished from the zero dose point. Measure 10 zero standard points at the same time to obtain the average value and standard deviation of RLU. Calculate the difference between the average value of RL...

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Abstract

The invention discloses a kit for early diagnosis of bladder cancer and a method for preparing the kit, and relates to the field of biomedical immune analysis. The detecting kit, which is fast, sensitive, highly specific in the early diagnosis of the bladder cancer is created by detecting a tumor marker of the bladder cancer tumor marker, namely, abnormally glycosylated integrin AG-alpha3beta, in human urine by means of microplate chemiluminescence immunoassay. The kit provided by the invention has the advantages of simplicity, convenience, quickness, sensitivity, high specificity, stability and the like, can be applied to early screening of the bladder cancer in a routine examination and to tracking observation and prognostic evaluation of the bladder cancer, and can satisfy the need that the existing in-vitro diagnosis of the bladder cancer is lack in specific diagnosis method.

Description

technical field [0001] The invention relates to a method for detecting tumor markers in the urine of bladder cancer patients. The method non-invasively realizes the early diagnosis of bladder cancer by quantitatively detecting the content of abnormally glycosylated integrin AG-α3β1 in the urine . Background technique [0002] Bladder cancer is the most common malignant tumor of the urinary system. In the United States, the incidence rate is listed as the fourth among male malignant tumors, the eighth among females, and the seventh among malignant tumors. In my country, the incidence rate of urological tumors ranks first. About 95% of bladder cancer originates from the bladder epithelium, most of which are malignant, and only a few are benign, including papilloma, transitional cell carcinoma, squamous cell carcinoma and adenocarcinoma. Among them, transitional cell carcinoma is the most common, accounting for more than 80%. At present, the curative effect of surgery, radio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N21/76
Inventor 范祖森杜颖李翀张倩云王彦英杨昭周鹏
Owner INSITUTE OF BIOPHYSICS CHINESE ACADEMY OF SCIENCES
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