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Method for detecting, identifying and/ or quantifying compound using adapter type reagent

An aptamer and compound technology, applied in the field of biomedical testing, can solve the problems affecting the flexibility of detection methods, affecting the accuracy and precision of detection, aptamer recognition affinity, specificity changes, etc. Short test time, good detection specificity and reproducibility

Inactive Publication Date: 2008-03-19
THE FIRST AFFILIATED HOSPITAL OF THIRD MILITARY MEDICAL UNIVERSITY OF PLA
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  • Summary
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AI Technical Summary

Problems solved by technology

In the existing technology of using aptamers to detect target molecules, it is necessary to directly connect the detection markers to the aptamers, which not only affects the flexibility of the detection method itself, but also affects the three-dimensional conformation of the aptamers due to the detection markers. , especially when the detection marker is relatively large, it will directly cause changes in the recognition of aptamers, the affinity and specificity of binding target molecules, thus affecting the accuracy and precision of detection

Method used

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Examples

Experimental program
Comparison scheme
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Embodiment Construction

[0036] Taking the preparation method of a high-affinity DNA aptamer capable of binding to human immunoglobulin E as an example, proceed as follows:

[0037] 1. Preparation and Purification of Proteins

[0038] The preparation and purification of the involved human immunoglobulin E are techniques known in the art (commercially available), and the pure human immunoglobulin E will be obtained as the screening target substance.

[0039] 2. Construction of random single-stranded DNA (ssDNA) library and primers

[0040] (1) Synthetic random ssDNA library

[0041] Artificially synthesized random ssDNA library with a length of 79 bp, a single-stranded DNA library with fixed sequences at both ends and random sequences in the middle 35 bases, with a storage capacity of 10 14-15 [5'-CCC CTG TCA GAA GTG TTT CTCAA-(N35)-AGA GAT GAG CAG GCG GAT ACT-3'], where N represents any one of the bases A, G, C, T. The random single-stranded DNA library was synthesized by Biosynthetics.

[0042] (...

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PUM

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Abstract

The present invention relates to a method using adapter type reagent to detect, identify, and / or ration the compound. The present invention adopts the SELEX technology, uses various biologic molecules possessing the diagnosing value including the nucleotide sequence, the protein, the aminophenol, the polypeptide chain, the glucide, or the complete thalli as the target matters, and filters the corresponding high specificity adapter from which to obtain the target matters. The present invention has the advantages that the repeatability and the stability of the test result can be improved greatly, lots difference of the test can be reduced, and the reliability of the test result can be fully ensured. The present invention can be used to diagnose the clinic target molecular quickly and simply, and can provide favorable basis for the lab diagnosis. The present invention has short test time, low reagent cost, short research and development period, stable quality, etc.

Description

technical field [0001] The invention belongs to the field of biomedical testing. The invention relates to a method for detecting, identifying and / or quantifying compounds using an aptamer-type reagent, and also relates to a preparation method for an aptamer-type detection kit used in the method. Background technique [0002] Most of the reagents currently used in clinical testing are antibodies, and most of them are imported reagents. Due to the differences and stability problems in the antibody preparation process, it is determined that a standard curve must be made for each batch of reagents in the quantitative determination, which makes the operation process complicated and cumbersome, and some substances with little or no immunogenicity cannot be detected. . Aiming at the above problems, the present invention adopts aptamer technology, and through the screening process of SELEX technology, obtains high-affinity specific aptamers of various target substances with clinica...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/53G01N33/531G01N33/546
Inventor 姚春艳府伟灵齐永志黄君富
Owner THE FIRST AFFILIATED HOSPITAL OF THIRD MILITARY MEDICAL UNIVERSITY OF PLA
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