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Chromatographic system for nucleic acid detection

A technology of nucleic acid and chromatographic medium, which is applied in the field of nucleic acid detection, and can solve problems such as a large number of samples, long hours of work, and insensitive reaction results.

Inactive Publication Date: 2012-05-23
BODITECHMED INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] However, the above-mentioned method is a complicated process in which both the preparation and reaction of the sample and the imaging require long hours of work
Moreover, the result of this reaction is not very sensitive, requires a large number of injections, and is very sensitive to different reaction conditions such as hybridization time or temperature, and reaction buffer solution or primers used.

Method used

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  • Chromatographic system for nucleic acid detection
  • Chromatographic system for nucleic acid detection
  • Chromatographic system for nucleic acid detection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0076] Preparation of Target Analytes and Capture Molecules

[0077] In order to prepare test strips for detecting HPV DNA, target analytes were prepared as follows. First, genomic DNA was extracted from a biopsy sample identified as HPV subtype 16, 18, or HPV positive (the biopsy sample was obtained from Yonsei University in Seoul, Korea, and its type was identified using a DNA chip). The extracted genomic DNA was then purified by phenol / chloroform extraction, in which 1 ml of the DNA solution was mixed with 1 ml of phenol / chloroform / isoamyl alcohol (25:24:1) followed by slow rotation and centrifuged at 14,000 rpm at room temperature 5 minutes. After centrifugation, transfer the supernatant to a clean test tube, add two volumes of 100% ethanol to it at -20°C and mix well. The supernatant of the mixture was then removed by centrifugation at 14,000 rpm for 15 min at 4°C, and the precipitate was gently washed with 70% EtOH. After centrifugation at 14,000 for 15 min at 4 °C to...

Embodiment 2

[0082] Pretreatment of sample pads

[0083] The flow rate of the components / analytes in the liquid sample is increased and the sensitivity is ensured by pre-treatment of the sample pad, and the non-specific binding of target analytes and / or captured DNA on the nitrocellulose membrane is reduced. Dip the sample pad (2.5x 30cm) into a solution containing 1% BSA, 0.05% Phosphate buffered saline (PBS, 137mM NaCl, 2.7mM KCl, 10mM disodium hydrogen phosphate, 2mM potassium dihydrogen phosphate, pH 7.4) for 10 minutes to reach equilibrium. The sample pads were then drained at 50°C to remove excess solution to prevent deformation. Store the pretreated sample pads under dry conditions until use.

Embodiment 3

[0085] Immobilization of capture sequences on nitrocellulose membranes

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Abstract

The present invention relates to a strip for detecting nucleic acids in analysis samples, based on the principles of lateral motion development technique and sequence-specific hybridization of nucleic acids, and to a nucleic acid detection system comprising the strip and also to a specific nucleic acid detection method employing the strip. The strip, system and method of the present invention allow not only a very high degree of ease in use but also of detection sensitivity and specificity, and allow accurate and rapid analysis even when a small amount of nucleic acid is employed.

Description

technical field [0001] The present invention mainly relates to the field of nucleic acid detection. In particular, the present invention relates to a test strip, method and system based on lateral flow chromatographic separation and sequence-specific detection of nucleic acids in samples. Background technique [0002] In research fields and industrial fields where nucleic acid detection is unavoidable, whether the detection involves confirmation of the presence or absence of target nucleic acid or qualitative or quantitative determination of nucleic acid, methods involving sequence-specific hybridization of nucleic acid are mainly used . One such method is the polymerase chain reaction-based test, in which specific target sequences are selectively amplified and the amplified products are analyzed by gel electrophoresis, followed by sequence-specific hybridization methods such as southern or northern blot or use dye staining for visualization. [0003] However, the above-m...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68G01N33/50G01N30/90
CPCG01N33/5308C12Q1/70G01N33/54386C12Q1/708C12Q2565/625C12Q2563/107C12Q1/6888G01N30/90
Inventor 崔义烈南基风郑东锡金成中
Owner BODITECHMED INC