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Kit and detection method for detecting protein expression of peripheral blood circulating tumor cell AR-V7 of prostate cancer patient

An AR-V7, peripheral blood circulation technology, applied in tumor/cancer cells, cell dissociation methods, animal cells, etc., can solve false negatives, false positives in detection methods, and tissue samples cannot evaluate the real-time dynamic status of AR-V7 in patients, etc. problems, to achieve the effect of reducing cell loss, good stability, and avoiding edge effects

Pending Publication Date: 2020-10-09
SHANDONG FIRST MEDICAL UNIV & SHANDONG ACADEMY OF MEDICAL SCI
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Problems solved by technology

[0007] Aiming at the disadvantages of detecting patients with advanced tumors or recurrent prostate cancer in real-time or repeated punctures to obtain tissue samples in the prior art, and evaluating the real-time dynamic status of AR-V7 in patients, and the existing detection methods are prone to false positives and false negatives, this paper The invention provides a kit and detection method for detecting the expression of AR-V7 protein in peripheral blood circulating tumor cells of prostate cancer patients: the circulating tumor cells (CTC) in the peripheral blood of patients with advanced prostate cancer are separated by using a membrane filtration device, and the immune group is further used to Detection of AR-V7 expression on CTC by chemical technology

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  • Kit and detection method for detecting protein expression of peripheral blood circulating tumor cell AR-V7 of prostate cancer patient
  • Kit and detection method for detecting protein expression of peripheral blood circulating tumor cell AR-V7 of prostate cancer patient
  • Kit and detection method for detecting protein expression of peripheral blood circulating tumor cell AR-V7 of prostate cancer patient

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Embodiment 1

[0048] 1. Use a membrane filtration device to separate and obtain CTCs in the peripheral blood of advanced or recurrent prostate cancer patients whose tissue samples cannot be obtained, and determine whether CTCs exist:

[0049] Collect 5ml of fasting blood from the median vein of the elbow for 8-12 hours, dilute the peripheral blood with 45ml of diluent, and then add 3ml of 4% paraformaldehyde to fix the diluted blood sample for 10 minutes;

[0050] At fixed intervals, assemble the membrane filtration device: as attached figure 1 , figure 2 , image 3 As shown, the filtering device is composed of a filter 3, a filter membrane 7, a blood sample container 2, a waste liquid tank 5, and an iron stand 1;

[0051] Wet the filter 3 with 10ml of PBS, and then add the fixed peripheral blood sample into the blood sample container 2 of the membrane filtration device, so that it is naturally filtered by gravity, and the CTC is trapped on the filter membrane 7;

[0052] The diameter o...

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Abstract

The invention provides a kit and a detection method for detecting protein expression of peripheral blood circulating tumor cells AR-V7 of a prostate cancer patient and. The kit comprises a diluent, adecolorizing solution, a staining solution A, a staining solution B, a mouse anti-human AR-V7 primary antibody, a horse radish peroxidase labeled goat anti-mouse secondary antibody, 0.1% Triton X-100,0.3% H2O2 and a reagent A, according to the detection method, a membrane filtration device is used for separation to obtain peripheral blood circulating tumor cells CTC, and an immunohistochemical technology is used for detecting the AR-V7 expression condition of the peripheral blood CTC. By utilizing the detection method provided by the invention, the AR-V7 expression condition of a patient withadvanced or recurrent hepatocellular carcinoma can be detected without acquiring a tissue specimen through needle biopsy, and real-time dynamic detection can be realized by utilizing a minimally invasive technology; by adopting the separation device and the diluent, the circulating tumor cells are well separated, the interference of blood cells is avoided, the false positive result caused by theedge effect possibly generated in the dyeing process is avoided, the stability is good, the cell loss is reduced, and the detection accuracy is improved.

Description

technical field [0001] The invention provides a kit and a detection method for detecting the expression of AR-V7 protein in peripheral blood circulating tumor cells of prostate cancer patients, and belongs to the technical field of molecular biology. Background technique [0002] Worldwide, the incidence of prostate cancer ranks second among male malignancies, and cancer-specific death ranks sixth. According to the 2015 China Cancer Registration Annual Report, the incidence of prostate cancer in China ranked 9th among malignant tumors in 2011, and the incidence and mortality of prostate cancer in urban men ranked 6th and 9th among male malignant tumors, respectively. The difficulty in the treatment of prostate cancer is metastatic prostate cancer, especially castration resistance after endocrine therapy. At present, the treatment methods for metastatic prostate cancer after castration resistance are limited, and all conventional treatment methods (radiotherapy, chemotherapy,...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/574C12N5/09
CPCG01N33/57484G01N33/57434C12N5/0694G01N2333/723C12N2509/10
Inventor 王焕昇李胜李浩高德海
Owner SHANDONG FIRST MEDICAL UNIV & SHANDONG ACADEMY OF MEDICAL SCI
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