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
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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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