Kit for detecting metastatic castration resistant prostate cancer (mCRPC) drug resistance

A castration-resistant, prostate cancer technology, applied in the field of immunoassay detection, can solve the problem of maintenance for 1.5 to 4 years, and achieve the effects of improved sensitivity, sensitive detection, and enhanced probe sensitivity

Active Publication Date: 2016-07-20
SHANGHAI MAJORBIO BIO PHARM TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although castration therapy has a certain effect, the control of the tumor can only be maintained for 1.5 to 4 years [Li Ming. Review of Prostate...

Method used

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  • Kit for detecting metastatic castration resistant prostate cancer (mCRPC) drug resistance
  • Kit for detecting metastatic castration resistant prostate cancer (mCRPC) drug resistance
  • Kit for detecting metastatic castration resistant prostate cancer (mCRPC) drug resistance

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037]Embodiment 1PSA-oligonucleotide probe

[0038] (1) Aldehyde modification of oligonucleotide probes

[0039] 50 nmol of Oligo1 (oligonucleotide) was prepared into a solution with 0.1 M pH 7.4 phosphate buffer. Weigh 500nmol of SFB (N-succinimidyl 4-formylbenzoate), dissolve it with anhydrous DMF (N,N-dimethylformamide), react at room temperature for 2.5h, and purify through a column to obtain Oligo -FB (aldehyde modified oligonucleotide).

[0040] Detection of the concentration of Oligo-FB: detection of A by Nanodrop spectrophotometer 260 The calculated value of Oligo-FB is 0.63nmol / μL.

[0041] Detection of aldehyde group modification rate: Use quantitative 2-hydrazinopyridine-2-hydrochloride solution to detect the aldehyde group modification rate. Take the above-mentioned Oligo-FB and add it to the 2-hydrazinopyridine-2-hydrochloride solution, shake and mix well, and react at 37°C for 1 hour. The absorbance value at 360nm detected by Nanodrop is 1.4, and the modific...

Embodiment 2

[0054] The QPCR amplification specific detection of embodiment 2Oligo molecule

[0055] Oligo1-13 molecules were diluted to three concentrations of 25000 molecules / μl, 83333 molecules / μl and 250000 molecules / μl. Then, taking Oligo1-3 as an example, mix Oligo1-3 at the same concentration to obtain mixed samples A, B, and C, as shown in Table 2.

[0056] Table 2 Example 2 sample formula

[0057]

[0058] The above samples were configured with QPCR amplification solution according to Table 3 to obtain a QPCR amplification kit, and then Oligo1-3 was subjected to QPCR amplification according to the procedures shown in Table 4, and Oligo1-3 in mixed samples A, B, and C Each was subjected to QPCR amplification, using the extended Oligo1-3 as a template, and the Ct values ​​of the amplification results are shown in Table 5. Among them, the extension primer is RT-P, its 3' end is complementary to the 3' end of the oligonucleotide, the 5' end can form a hairpin structure, and the m...

Embodiment 3

[0072] Embodiment 3 makes standard curve

[0073] The same result can be obtained with Oligo1-13. In order to simplify the process, the following examples use Oligo1-3 as an example.

[0074] According to the steps of Example 1, Oligo2 was modified with SFB having a molar equivalent of 5 times to obtain Oligo-FB, and PSMA was modified with a hydrazine group of SANH having a molar equivalent of 10 times to obtain PSMA-SANH, and the molar ratio was 10:1 PSMA-Oligo2 was obtained after reacting Oligo-FB with PSMA-SANH at room temperature for 16 hours.

[0075] According to the steps of Example 1, Oligo3 was modified with SFB with a molar equivalent of 20 times to obtain Oligo-FB, and PSMA was modified with hydrazine groups with SANH with a molar equivalent of 50 times to obtain PSMA-SANH, and the molar ratio was 8:1 PSMA-Oligo3 was obtained after reacting Oligo-FB with PSMA-SANH at room temperature for 24 hours.

[0076] PSA-Oligo1, PSMA-Oligo2 and PSMA-Oligo3 were subjected to ...

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Abstract

The invention provides a kit for detecting metastatic castration resistant prostate cancer (mCRPC) drug resistance. The kit is characterized by comprising an antibody-oligonucleotide probe obtained by independently coupling PSA (prostate specific antigen) and PSMA (prostate specific membrane antigen) antibodies with different oligonucleotides; an amplification primer and a buffer solution for performing PCR (polymerase chain reaction) amplification on the antibody-oligonucleotide probe; and a fluorescent probe. By detecting the expression quantity of PSA and PSMA antigens, the kit can analyze the relative activity of AR signals, thereby detecting the mCRPC drug resistance of a patient.

Description

technical field [0001] The invention relates to the field of immunoassay detection, in particular to a kit for detecting drug resistance of metastatic castration-resistant prostate cancer. Background technique [0002] Prostate cancer is one of the most common tumors in men. It is not easy to be found in the early stage, and the disease is usually developed to an advanced stage when it is diagnosed. The mainstay of treatment for advanced prostate cancer is endocrine therapy. In prostate cancer, androgens can promote tumor growth, so androgen deprivation therapy is used to reduce androgen levels to achieve "castration". Although castration therapy has a certain effect, the control of the tumor can only be maintained for 1.5 to 4 years [Li Ming. Review of Prostate Hot Issues. Modern Journal of Urogenital Tumors. 3.3 (2011): 129-131], and further development will follow For castration-resistant prostate cancer (CRPC). Even metastases to other organs other than the prostate, ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6804C12Q1/6886C12Q2600/106
Inventor 陈昌岳李静蔡红东邓文斌甘广利张祥林
Owner SHANGHAI MAJORBIO BIO PHARM TECH
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