Nucleic acid aptamer of cortisol, and gold-colloidal test strip used for detecting cortisol

A nucleic acid aptamer, gold-labeled test paper technology, applied in measurement devices, instruments, scientific instruments, etc., can solve the problems of expensive instruments, time-consuming, complicated processes, etc., and achieve a simple detection method, a wide range of applications, and good stability. Effect

Inactive Publication Date: 2017-11-17
华测康诺(武汉)生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The results of various commercial immunoassay kits are highly variable, and the cost of the kits is also relatively expensive
Although the LC-MS method is accurate and sensitive, it requires expensive instruments, so it is difficult to popularize in general laboratories
Various traditional detection methods are complex, time-consuming, costly, and cannot guarantee batch-to-batch stability

Method used

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  • Nucleic acid aptamer of cortisol, and gold-colloidal test strip used for detecting cortisol
  • Nucleic acid aptamer of cortisol, and gold-colloidal test strip used for detecting cortisol
  • Nucleic acid aptamer of cortisol, and gold-colloidal test strip used for detecting cortisol

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] (1) Synthesis of nucleic acid aptamers and complementary probes

[0031] Synthesis of thiol-modified aptamers at the 5' end:

[0032]5'-SH-GCAGAGGATCATGATGCGGGGTGGAGAATGGGCCGCACTTCGGCTTCACTCTTGGCAGAAAAGCTT-3';

[0033] Synthesis of complementary probe A modified with biotin at the 5' end:

[0034] 5'-biotin-AAGCTTTTCTGCCAAGAGTGAAGCCGAAGT-3'.

[0035] The following are the nucleic acid aptamers and corresponding probes used in the comparison test:

[0036] Synthesis of thiol-modified aptamers at the 5' end for comparison:

[0037] 5'-SH-CAACGAGTCACTGTTTCTGGGTGGAGAATGGGCCGCACTTCGGCTTCACAACGAGTCACTGTTTCT-3';

[0038] Complementary probes synthesized with biotin-modified 5' ends of the alignment:

[0039] 5'-biotin-AGAAACAGTGACTCGTTGTGAAGCCGAAGT-3'.

[0040] (2) Preparation of colloidal gold solution

[0041] 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, and then add 9 μL of 100 μM sul...

Embodiment 2

[0056] Gold standard test strips were prepared and used according to the method described in Example 1. Take saliva samples containing 0nM, 10nM, 15nM, 20nM, 50nM cortisol respectively for detection, and the detection results are as follows: Figure 4 As shown on the left side of A, the T-line color development is from strong to weak, and the detection concentration range is 0 to 50nM; urine samples containing 0nM, 50nM, 100nM, 150nM, and 200nM cortisol were taken for detection, and the detection results are as follows Figure 4 As shown on the right side of A, the T-line color development ranges from strong to weak, and the detection concentration range: 0-200nM.

[0057] Figure 4 B is the result of the comparison test (using the comparison nucleic acid aptamer and the comparison probe in Example 1), get the saliva samples containing respectively 0nM, 10nM, 15nM, 20nM, 50nM cortisol for detection, and the detection results are as follows Figure 4 As shown on the left side...

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Abstract

The invention discloses a cortisol nucleic acid aptamer and a gold standard test strip for detecting cortisol, which belong to the technical field of hormone detection. The sequence of the nucleic acid aptamer of the present invention is shown in SEQ ID NO.1, and the sequence of its complementary probe A is shown in SEQ ID NO.2. The gold standard test strip of the invention comprises a sample glass fiber membrane, a nucleic acid probe gold standard pad, a nitrocellulose membrane, a water-absorbing glass fiber membrane and a plastic liner. The nucleic acid aptamer of cortisol labeled with colloidal gold is attached to the nucleic acid probe gold standard pad; the detection line and the reference line are drawn on the nitrocellulose membrane, the detection line is drawn by the cortisol-bovine serum albumin conjugate solution, and the reference line is drawn by A solution of biotin-modified complementary probe A conjugated with streptavidin was drawn. The invention does not need other auxiliary equipment, can complete the detection of cortisol within 10 to 15 minutes, and the result is accurate and reliable; the sensitivity and specificity are high, the stability is good, the cost is low, and the application range is wide.

Description

technical field [0001] The invention belongs to the technical field of hormone detection, and in particular relates to a cortisol nucleic acid aptamer and a gold standard test strip for detecting cortisol. Background technique [0002] Cortisol is a hormone called a corticosteroid that can be produced naturally by the body. It’s also called the stress hormone because humans produce more of it when they’re under significant stress. Most noticeable during counterreaction. The adrenal glands are responsible for producing cortisol, and the amount produced varies throughout the day, with the body tending to use more cortisol in the morning. The level of cortisol rises sharply within 30 minutes after getting up every morning, and reaches the highest value about 30 to 45 minutes after getting up, and then gradually decreases throughout the day until reaching the lowest value before going to bed. [0003] The normal content of cortisol in the human body can ensure that the requir...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/531G01N33/559
CPCG01N33/531G01N33/559
Inventor 程克文李斌
Owner 华测康诺(武汉)生物科技有限公司
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