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Gold-labeled strip based on nucleic acid aptamers of progesterone in saliva and used for detection

A nucleic acid aptamer and gold standard test paper technology, which is applied in biological testing, measuring devices, genetic engineering, etc., can solve problems such as inability to guarantee batch-to-batch stability, high operational technical requirements, and complicated processes, and achieve simple sampling and application The effect of wide range and simple detection method

Inactive Publication Date: 2016-06-08
李斌
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

As a standard method for detecting progesterone in serum, chromatography has incomparable advantages in terms of accuracy, selectivity, reproducibility, and sensitivity, but it also has some limitations: such as expensive instruments, high technical requirements for operation, Requires complicated sample pretreatment, etc.
[0006] Various traditional detection methods are complex, time-consuming, costly, and cannot guarantee batch-to-batch stability

Method used

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  • Gold-labeled strip based on nucleic acid aptamers of progesterone in saliva and used for detection
  • Gold-labeled strip based on nucleic acid aptamers of progesterone in saliva and used for detection
  • Gold-labeled strip based on nucleic acid aptamers of progesterone in saliva and used for detection

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Embodiment 1 A kind of preparation of gold standard test paper strip based on progesterone nucleic acid probe

[0038] (1) Preparation of colloidal gold solution

[0039] 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 for 1 hour , to obtain the nucleic acid aptamer solution with activated sulfhydryl groups.

[0040] Add 100 μL of 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 the colloidal gold intermediate solution 1 modified by the nucleic acid aptamer.

[0041] 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 ...

Embodiment 2

[0053] 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.

[0054] figure 2 It is a schematic diagram of the basis for judging the detection results during the detection process of progesterone 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:

[0055] figure 2 The left one in the cente...

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Abstract

The invention discloses a gold-labeled strip based on nucleic acid aptamers of progesterone in saliva and used for detection. The 5' end of a sequence of the nucleic acid aptamers of progesterone is modified by sulfydryl, the 5' end of a complementary probe A sequence is modified by biotin, and the 5' end of a complementary probe B sequence is modified by biotin. According to the gold-labeled strip for detecting progesterone, a sample glass fiber membrane, a nucleic acid probe gold-labeled cushion, a nitrocellulose membrane and a water absorption glass fiber membrane are sequentially arranged on a plastic substrate in a lap joint mode, a detection line and a reference line are drawn on the nitrocellulose membrane, and the reference line is close to one end of the water absorption glass fiber membrane. Colloidal gold with the surface modified by the nucleic acid aptamers is attached to the nucleic acid probe gold-labeled cushion, the detection line is drawn with a nucleic acid aptamer A solution, and the reference line is drawn with a nucleic acid aptamer B solution. No other auxiliary instrument or equipment is needed, detection can be completed within 10-15 min, and the result is accurate and reliable; sensitivity and specificity are high, stability is good, cost is low, and the application range is wide.

Description

technical field [0001] The invention relates to the technical field of hormone detection, in particular to a nucleic acid aptamer based on progesterone in saliva and a gold standard test strip for detection. Background technique [0002] Progesterone, also known as progesterone, is a natural progesterone produced by the ovary and placenta with a molecular weight of 314.46. It works with estrogen to maintain the female reproductive cycle and female physiological characteristics. Dynamic monitoring of progesterone content in serum is helpful to determine ovulation period, understand luteal phase function, and study the mechanism of action of various steroid contraceptives and anti-early pregnancy drugs. In addition, studies in recent years have shown that the role of progesterone has gone far beyond the scope of reproductive function. It can not only be synthesized and secreted in the nervous system, but also affect the structure and function of the nervous system. [0003] P...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/115G01N33/74G01N33/531
CPCC12N15/115C12N2310/16G01N33/531G01N33/74G01N2333/575
Inventor 彭杉
Owner 李斌
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