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A nucleic acid aptamer based on estradiol in saliva and a gold standard test strip for detection

A nucleic acid aptamer, gold-labeled test paper technology, applied in biological testing, measuring devices, material inspection products, etc., can solve problems such as inability to guarantee inter-batch stability, complex testing process, and long time-consuming, and achieve simple sampling and detection. Simple method and low cost effect

Inactive Publication Date: 2017-11-17
李斌
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The detection of changes in the level of estradiol in the body has very important clinical significance. However, the traditional detection methods are generally based on urine or blood, which brings inconvenience or pain to users during the sampling process. , and the detection process is complex, time-consuming, costly, and the stability between batches cannot be guaranteed

Method used

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  • A nucleic acid aptamer based on estradiol in saliva and a gold standard test strip for detection
  • A nucleic acid aptamer based on estradiol in saliva and a gold standard test strip for detection
  • A nucleic acid aptamer based on estradiol in saliva and a gold standard test strip for detection

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Embodiment 1

[0036] Embodiment 1 A kind of preparation of gold standard test paper strip based on estradiol nucleic acid probe

[0037] (1) Preparation of colloidal gold solution

[0038]Mix 1 μL of 0.5M acetic acid buffer solution with a pH of 5.9 and 1.5 μL of 10 mM TCEP (tris(2-carboxyethyl)phosphine) solution, then add 9 μL of 100 μM nucleic acid aptamer solution, mix well and react at 25°C After 1 hour, the nucleic acid aptamer solution with activated sulfhydryl group was obtained.

[0039] Add 100 μL of a 30-40 nm gold nanoparticle solution to the nucleic acid aptamer solution with activated sulfhydryl groups, and shake at 25° C. for 30 minutes to obtain a nucleic acid aptamer-modified colloidal gold intermediate solution 1 .

[0040] Add 85 μL of 10 μM dATP solution to the aptamer-modified colloidal gold intermediate solution 1, shake and react at 25°C for 15 minutes, then let stand for 30 minutes, and then stand at 4°C for 6 hours to obtain aptamer-modified Colloidal gold interme...

Embodiment 2

[0052] see figure 2 , where No. 1 is the sample glass fiber membrane with a length of 18mm; No. 2 is the nucleic acid probe gold label pad with a length of 4mm; No. 3 is a nitrocellulose membrane with a length of 20mm; No. 6 is a water-absorbing glass fiber membrane with a length of 18mm , No. 4 is the detection line with a width of 1 mm, and No. 5 is the reference line with a width of 1 mm. The arrows in the figure indicate the direction of liquid capillary action.

[0053] figure 2 It is a schematic diagram of the basis for judging the detection results during the detection process of estradiol in saliva by using the gold standard test strip of the present invention. How to judge the content of the target substance in the test sample through the test results of the gold standard test strip based on the nucleic acid probe is an important part of the test method. The specific test results are judged based on the following:

[0054] figure 2 The left one in the center i...

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Abstract

The invention discloses a nucleic acid aptamer based on estradiol in saliva and a gold mark test strip for detection. The 5' terminal of the sequence of the nucleic acid aptamer based on estradiol is modified with sulfydryl, the 5' terminal of the sequence of a complementary probe A is modified with biotin, and the 5' terminal of the sequence of a complementary probe B is modified with biotin. The gold mark test strip for detecting estradiol is formed by arranging a sample glass fiber membrane, a nucleic acid probe gold mark pad, a nitrocellulose membrane and a water-absorbing glass fiber membrane on a plastic lining plate in sequence in a lap joint mode, wherein the nitrocellulose membrane is marked with a detection line and a reference line close to one end of the water-absorbing glass fiber membrane, colloidal gold with the surface modified with the nucleic acid aptamer is attached to the nucleic acid probe gold mark pad, the detection line is drawn with a nucleic acid aptamer solution A, and the reference line is drawn with a nucleic acid aptamer solution B. By means of the nucleic acid aptamer based on estradiol in saliva and the gold mark test strip for detection, detection can be completed within 10-15 without other secondary instruments, and results are accurate and reliable; the nucleic acid aptamer based on estradiol in saliva and the gold mark test strip for detection are high in sensitivity and specificity, good in stability, low in cost and wide in application range.

Description

technical field [0001] The invention relates to the technical field of hormone detection, in particular to a nucleic acid aptamer based on estradiol in saliva and a gold standard test strip for detection. Background technique [0002] Estradiol (17β-estradiol) is an endogenous estrogen with a molecular weight of 272.38. It is the strongest biological activity expressed by various hormones in women. With the in-depth research on estradiol, it is found that the metabolites produced by endogenous estrogen can still exert various biological effects, and they do not have carcinogenic effects under normal circumstances. The ovaries secrete estrogen, and diseases such as anovulatory hemorrhage, polycystic ovary syndrome, and functional ovarian tumors can lead to increased secretion of estrogen by the ovary. Intimal hyperplasia and canceration. [0003] With the development of society, emotional diseases are getting more and more attention from the society, and women are more pron...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/115G01N33/74G01N33/558
CPCG01N33/558G01N33/74
Inventor 彭杉
Owner 李斌
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