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Detection kit taking AKR1C1 as detection target and use method of detection kit

A technology for detecting kits and targets, which is applied in the field of biomedicine, can solve the problems of detection kits and AKR1C1-specific primer design difficulties, and achieve high accuracy

Pending Publication Date: 2022-01-04
SHANGHAI FIRST PEOPLES HOSPITAL +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These highly homologous family factors add difficulty to the design of AKR1C1-specific primers, and also bring difficulties to the development of detection kits with AKR1C1 as the detection target

Method used

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  • Detection kit taking AKR1C1 as detection target and use method of detection kit
  • Detection kit taking AKR1C1 as detection target and use method of detection kit
  • Detection kit taking AKR1C1 as detection target and use method of detection kit

Examples

Experimental program
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Effect test

Embodiment 1

[0027] This embodiment provides a detection kit using AKR1C1 as a detection target, and the detection kit includes:

[0028] 1. Primers

[0029] The mRNA sequences of AKR1C1, AKR1C2, and AKR1C3 were downloaded from the GenBank library, and compared with Vector NTI software, it was found that the CDS regions of the three proteins of the AKR1C family were highly homologous. Primers were designed using snapgene software, and verified by NCBI BLAST comparison, the designed primers were indeed only specifically matched to AKR1C1 mRNA (NM_001353.6). The size of the amplified fragment of the primers is 193bp, and the sequences of the primers are as follows.

[0030] Amplification Primer 1:

[0031] Upstream primer: 5'-AGGTCGGGGCATGGAAAAAT-3' (SEQ ID NO.1);

[0032] Downstream primer: 5'-CGCATTCTCAGGAGACCGTT-3' (SEQ ID NO.2).

[0033] 2. Reagents

[0034] The above-mentioned amplification primer 1 is placed in the kit, and then the following reagents are loaded into the kit:

[...

Embodiment 2

[0038] This embodiment provides a detection kit using AKR1C1 as the detection target. The difference between the detection kit and the detection kit in Example 1 is that the primers of the detection kit include: amplification primer 2, the amplification primer 2 The amplification primer 2 adopts the same primer design method as in Example 1, the size of the amplified fragment is 141bp, and the primer sequence is as follows:

[0039] Upstream primer: 5'-CAGGTGTCTCACGACCTGAC-3' (SEQ ID NO.3);

[0040] Downstream primer: 5'-GAAGTAGTGGGCCGCTTACA-3' (SEQ ID NO.4).

Embodiment 3

[0042] This embodiment provides a detection kit using AKR1C1 as the detection target. The difference between the detection kit and the detection kit in Example 1 is that the primers of the detection kit include: amplification primer 1 and amplification primer 3. The amplification primer 3 is used to amplify the internal reference gene GAPDH (Gene ID: 2597) to detect the relative expression level of AKR1C1. The size of the amplified fragment of amplification primer 3 is 208bp, and the primer sequence is as follows:

[0043] Upstream primer: 5'-AACGGATTTGGTCGTATTG-3' (SEQ ID NO.5);

[0044] Downstream primer: 5'-GGAAGATGGTGATGGGATT-3' (SEQ ID NO.6).

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Abstract

The invention discloses a detection kit taking AKR1C1 as a detection target and a use method of the detection kit. The detection kit comprises a specific primer pair used for detecting AKR1C1 expression. The specific primer pair comprises an amplification primer 1 and / or an amplification primer 2. The nucleotide sequence of an upstream primer of the amplification primer 1 is as shown in SEQ ID NO.1, and the nucleotide sequence of a downstream primer of the amplification primer 1 is as shown in SEQ ID NO.2. The nucleotide sequence of an upstream primer of the amplification primer 2 is as shown in SEQ ID NO.3, and the nucleotide sequence of a downstream primer of the amplification primer 2 is as shown in SEQ ID NO.4. The invention provides the detection kit taking AKR1C1 as the detection target for the first time, and the kit can specifically detect the expression of the AKR1C1 gene, assess whether a patient is resistant to progestational hormone or not in the early stage of the disease of the endometrial cancer patient, and further can be used for judging the applicability of the patient to progestational hormone treatment.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a detection kit using AKR1C1 as a detection target and a use method thereof. Background technique [0002] Endometrial cancer is one of the most common tumors of the female reproductive system, and its incidence is increasing year by year. It has become a malignant tumor of the reproductive tract that seriously threatens women's health. Studies have shown that the occurrence of endometrial cancer may be caused by the long-term stimulation of endometrium by estrogen without the antagonism of progesterone. At present, there are many treatment methods for endometrial cancer, such as surgery, radiotherapy, chemotherapy and so on. However, with the increase in the incidence of endometrial cancer and the improvement of diagnosis, many young patients need conservative treatment to preserve their reproductive function. Progesterone, as one of the means of conservative treatment, has achieved ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12Q1/6851C12N15/11
CPCC12Q1/6886C12Q1/6851C12Q2600/106C12Q2600/158C12Q2600/166C12Q2531/113C12Q2521/107C12Q2545/101C12Q2563/107
Inventor 张箴波覃祚树陈晓军宁程程
Owner SHANGHAI FIRST PEOPLES HOSPITAL
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