Bladder cancer detection kit

A detection kit and bladder cancer technology, applied in the field of molecular biology, can solve the problems of lowering the specificity of FISH detection, dependence on detection effect, and long time-consuming

Pending Publication Date: 2021-03-23
SUREXAM BIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The detection effect depends on the quality of the prepared probes, and the fluorescent in situ hybridization probes prepared by bacterial artificial chromosomes (BAC) etc. have many repetitive sequences that lead to the generation of non-specific fluorescent signals, which reduces the detection efficiency of FISH to a certain extent. specificity
Moreover, the existing FISH detection process takes a long time, and the hybridization time basically takes more than ten hours, resulting in high time cost for FISH detection

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Probe composition preparation in the bladder cancer detection kit of embodiment 1

[0057] In this embodiment, a bladder cancer detection kit includes detection probes for chromosomes 3, 7, and 17 and the p16 gene; the detection probes for chromosomes 3, 7, and 17 and the p16 gene include probe A respectively , B1 probe, B2 probe, C probe, D probe and E probe. When the detection probe described in this embodiment is in use, the detection probe for chromosome 3 and chromosome 7 is hybridized with the same sample to be detected, which is denoted as CEN3 / CEN7 probe; the detection probe for chromosome 17 and p16 gene The needle is hybridized with the same sample to be detected, which is recorded as CEN17 / p16 probe. in,

[0058] 1. A probe

[0059] The base composition of the A probe from the 5' end to the 3' end is as follows: P1 sequence, M1, specific P2 sequence, M1, P1 sequence.

[0060] (1) Specific P2 sequence

[0061] The specific P2 sequence is selected by the i...

Embodiment 2

[0093] Example 2 Using the kit in Example 1 to detect the sample

[0094] The formulations of the various solutions are shown in Table 9.

[0095] Formulation of various solutions of table 9

[0096]

[0097] In this embodiment, urine samples from patients with bladder cancer are preferred, and abnormalities of chromosomes 3, 7, and 17 and p16 gene in the exfoliated cells are detected, and the CEN3 / CEN7 probe mixture and the CEN17 / p16 probe mixture are all used in the embodiment 1 All probes in the corresponding list of bladder cancer detection kit.

[0098] 1. Pretreatment of samples to be tested

[0099] (1) Collect the sample to be tested: collect 40ml of the patient’s urine, divide it into two equal portions of 20ml each, and centrifuge to get the cell pellet. After washing with PBS, add 4ml of PBS to resuspend the cell pellet, add 1ml of fixative, and vortex Mix well, let stand at room temperature for 8 minutes, add the above two cell suspensions to two filter membr...

Embodiment 3

[0129] Embodiment 3 The establishment of the abnormal threshold value of the kit of the present invention

[0130] In order to establish the threshold value of the kit of the present invention for detecting abnormalities of chromosomes 3, 7, and 17 and the p16 gene, the following experiments were designed: 20 normal human urine specimens were collected, and the bladder cancer detection kit described in Example 1 of the present invention was used for detection . Each group of probes observed 50 cells, counted the percentages of different types of abnormal cells, and calculated the abnormal threshold according to the formula "abnormal threshold = mean (M) + 3 x standard deviation (SD)".

[0131] Using the kit prepared by the above design, according to the above experimental design, according to the detection process and method described in Example 2, 20 cases of normal human urine specimens (numbered 1 to 20) were detected, and each sample and each group of probes were read. Ne...

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Abstract

The invention provides a bladder cancer detection kit. The bladder cancer detection kit comprises at least one of detection probes for chromosomes 3, 7 and 17 and a gene p16; the detection probes forthe chromosomes 3, 7 and 17 and the gene p16 include a probe A, a probe B1, a probe B2, a probe C, a probe D and a probe E; a base of the probe A from the 5' end to the 3' end is composed of a P1 sequence, M1, a specific P2 sequence, M1 and a P1 sequence; the probe B1 is a P3 sequence; the probe B2 is a P4 sequence; a base of the probe C from the 5' end to the 3' end is composed of a P5 sequence,M2 and a P6 sequence; the probe D is a P7 sequence; and a base of the probe E from the 5' end to the 3' end is composed of a P8 sequence of which the 5' end is modified with a fluorophore and the 3' end is modified with a structure capable of quenching the fluorophore. The bladder cancer detection kit has the advantages of short hybridization time, high sensitivity, strong specificity, high accuracy, simple operation steps and the like.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and in particular relates to a bladder cancer detection kit. Background technique [0002] Bladder cancer is the most common malignant tumor of the urinary system, among which urothelial carcinoma accounts for about 90% of bladder cancer, and 75% of newly diagnosed patients are non-invasive bladder cancer. The postoperative recurrence rate of bladder cancer is as high as 50% to 70%, and 10% to 15% of recurrent non-invasive bladder cancer will progress to invasive or metastatic cancer (Kaufman D S et al., Lancet, 2009, 374:239 -249). Therefore, early diagnosis of bladder cancer and prevention of tumor recurrence are very important. Currently, cystoscopy and cytological examination are two methods commonly used in the clinical diagnosis of bladder cancer. However, cystoscopy is an invasive examination, which will bring some pain to patients, and it is not easy to detect early tumor les...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12N15/11
CPCC12Q1/6886C12Q2600/156
Inventor 吴诗扬彭璨璨许嘉森刘志明刘芳
Owner SUREXAM BIO TECH
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