Detection method for detecting prostate cancer circulating tumor cells

A technology for prostate cancer and tumor cells, which is applied in the preparation of test samples, measuring devices, sampling, etc., can solve problems such as no detection methods, and achieve good results.

Inactive Publication Date: 2017-04-26
上海市质子重离子医院有限公司 +2
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With the deepening of the research, the results show that PSMA is expected to become a tumor marker for pr...
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Abstract

The invention discloses a detection method for detecting prostate cancer circulating tumor cells, wherein the detection method comprises the following specific steps: step one, collection of a peripheral blood sample; step two, blood cell separation; step three, antibody staining; step four, cell immobilization; step five, ethanol dehydration; step six, probe incubation; step seven, cleaning; and step eight, sheet sealing. The method can be used for early detection of prostate cancer, so as to help diagnosis and treatment; the method can specifically detect free tumor cells in a circulating blood system, calculates out the positive rate, and guides doctors to select a treatment scheme according to the positive rate and the cell number condition of patients at different time nodes of a treatment course; the method can perform subsequent detection work, such as single cell sequencing, targeted drug screening and the like, detects other tumor substances simultaneously, is more comprehensive in detection of tumor metastasis, drug resistance, differentiation and other conditions, and has good use effect.

Application Domain

Preparing sample for investigation

Technology Topic

Circulating tumor cellNODAL +19

Examples

  • Experimental program(2)

Example Embodiment

[0020] Example 1
[0021] The LNCaP human prostate cancer cells were stained with antibodies according to the above method. The antibodies were prepared in antibody diluent. The concentration of CD45-Alexa 594 antibody and PSMA-Alexa 488 antibody were both 1mg/ml. Take 1μl of each and add 198μl of antibody diluent. . Read the film manually at room temperature. The equipment used includes a low-speed large-capacity centrifuge, a miniature desktop vacuum pump, a miniature horizontal decoloring shaker, a balance balance, an electric drying oven, a digital display constant temperature water bath, a 50 ml large magnetic stand, and a manual single channel Adjustable pipettes, micro-hair dryers, large-capacity electric pipettes, micro-centrifuge, vortex mixer, in-situ hybridization instrument and fluorescence microscope, free scanning software of Nikon microscope for software and NIS-Elements F 4.00.00 Take pictures.
[0022] The most critical criteria for CTC are CD45 negative and non-diploid chromosome 8. CD45: CTC sometimes sees a light red background in the red channel. The most obvious difference from white blood cells is the bright red halo that does not surround the nucleus (sometimes the time circle is incomplete).
[0023] CEP8: Monomer, diploid or multimer, the signal is very clear in the red or orange channel. Tumor marker channel: green signal, according to the expression of tumor marker, the signal is different in light and dark, and some tumor cells do not express. DAPI: blue, round or oval
[0024] CTC judgment:
[0025] DAPI+ / CD 45-/Tumor Mark+ / Chromosome 8+ (any number of chromosomes).
[0026] DAPI+ / CD 45-/Tumor mark-/ Chromosome 8+ (1 body, 3 body, 4 body, ≥5 body), due to the degradation of some tumor markers during the interstitial process of CTC, this kind of CTC is more common. CTC in peripheral blood can be divided into single CTC and tumor thrombus (CTM, ≥2 cancer cells). Cancer thrombus cells play a particularly critical role in the process of tumor metastasis and recurrence. Such cells cannot be ignored.

Example Embodiment

[0027] Example 2
[0028] The patient's peripheral blood CTC test results and related medical history are as follows: patient, male, 73 years old. The patient underwent radical prostatectomy for prostate cancer 5 years ago, and 2 years later due to local recurrent photon radiotherapy in the prostate fossa, with a total dose of 68Gy/34Fx. After radiotherapy, endocrine therapy and chemotherapy were given successively, but PSA was progressively increased, 2016.2 .26 line SPECT (99mTc-PSMA) systemic examination, suggesting: after prostate cancer, left iliac vessels, retroperitoneum multiple lymph nodes enlargement, radioactive uptake increased, consider metastasis; 2016.3.3 line based on prostate specific membrane antigen (PSMA) ) Is a negative enrichment method for molecular markers to detect circulating tumor cells in peripheral blood. The results show that: in 7.5ml of peripheral blood, a total of 15 CTCs were detected, of which 3 were PSMA+ CTCs (respectively 3 body +, 4 body +, 5+), the detection effect is real and effective.

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Description & Claims & Application Information

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