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Solid-phase particle material for treating biological sample and biological sample treatment method

A solid-phase particle, biological sample technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve the problems of time-consuming extraction steps, high reagent costs, complex steps, etc., and achieve high-throughput automated sample processing, The effect of saving materials and operating time, and inhibiting nuclease activity

Pending Publication Date: 2021-11-02
YEASEN BIOTECHNOLOGY (SHANGHAI) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The premise of nucleic acid detection is to obtain relatively pure nucleic acid from the sample to be tested, and the traditional nucleic acid extraction steps are usually time-consuming. Although automated nucleic acid extraction has been widely used, it still takes a long time, usually more than 30 minutes , and the steps are more complicated, and the reagent cost is relatively high
It is a simpler method to quickly release the nucleic acid in the biological sample and directly use it for PCR amplification, but the problem with this method is that the biological sample contains a variety of inhibitory components, which leads to poor PCR amplification effect and inaccurate detection results. Accurate, difficult for clinical application

Method used

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  • Solid-phase particle material for treating biological sample and biological sample treatment method
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  • Solid-phase particle material for treating biological sample and biological sample treatment method

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Effect test

Embodiment 1

[0032] The preparation of embodiment 1 agarose solid-phase granular material

[0033] Take 100 mL of suspension of magnetic agarose carboxyl microbeads (purchased from Yingruicheng Biochemical Technology Co., Ltd.), suck off the supernatant, add 100 mL of 50 mM MES buffer (pH 5.0), stir well to suspend the microbeads. Continue to add 1 g of 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride and 1 g of N-hydroxysuccinimide to the above solution, and stir thoroughly for 30 min. After the stirring, remove the reaction solution by magnetic suction and wash with water, continue to add 100 mL of MES buffer solution containing 1 g of iminodiacetic acid to the micro-beads, and stir thoroughly for 5 h. After the reaction is finished, the microbeads are washed with pure water to obtain the solid-phase granular material.

Embodiment 2

[0034] The preparation of embodiment 2 polystyrene carboxyl microbeads

[0035] Add 5 mL of styrene to a mixed solution of 70 mL of ethanol and 30 mL of water, then add 0.1 g of azobisisobutyronitrile, stir well, then raise the temperature to 70°C, and stir for 3 hours. After the reaction, the polystyrene microspheres were obtained by high-speed centrifugation, and washed several times with ethanol. Take 2 g of the prepared polystyrene microspheres, disperse them in 50 mL of tetrahydrofuran solution, add 1 g of succinic anhydride, 0.05 g of aluminum trichloride, stir and heat to 60 ° C for 9 h. After the reaction, the polystyrene carboxyl microbeads were obtained by high-speed centrifugation, and washed several times with ethanol.

Embodiment 3

[0036] The preparation of embodiment 3 polystyrene solid phase particle material

[0037] Take 100 mL of polystyrene carboxy microbead suspension, suck off the supernatant, and add 100 mL of 50 mM MES buffer (pH 5.0), stir well to suspend the microbeads. Continue to add 1 g of 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride and 1 g of N-hydroxysuccinimide to the above solution, and stir thoroughly for 30 min. After the stirring was completed, the reaction solution was removed by centrifugation, and 100 mL of MES buffer solution containing 1 g of iminodiacetic acid was added to the microbeads, and stirred thoroughly for 5 h. After the reaction is finished, the microbeads are washed with pure water to obtain the solid-phase granular material.

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Abstract

The invention provides a solid-phase particle material for treating a biological sample, the solid-phase particle material takes a high-molecular polymeric material or a gel material as a core, magnetic particles can be contained in the core or on the surface of the core, a composite structure of the magnetic particles and the core is externally coated with a polymer coating, the surface of the coating is modified with weak cation exchange groups, the weak cation exchange groups can chelate metal ions and adsorb proteins. The invention further discloses a corresponding biological sample treatment method. The related raw materials comprise the solid-phase particle material and the nucleic acid releasing agent. The solid-phase particle material is a weak cation exchange material, and the nucleic acid releasing agent has the effects of splitting cells and viruses, inhibiting nucleic acid degradation and the like. According to the reagent and the method provided by the invention, the sample can be simply and quickly treated, nucleic acid is released, part of impurities are removed, a nucleic acid extraction and purification step is not needed, the time and the material cost are greatly saved, and the reagent and the method have a relatively good application value.

Description

technical field [0001] The patent of the present invention relates to a solid-phase granular material for processing biological samples and a biological sample processing method, belonging to the field of biotechnology. Background technique [0002] PCR (Polymerase Chain Reaction, Polymerase Chain Reaction) is a molecular biology technique used to amplify and amplify target DNA fragments in vitro. It is a rapid detection method with high sensitivity and high specificity and has been widely used in Clinical testing and scientific research fields. The premise of nucleic acid detection is to obtain relatively pure nucleic acid from the sample to be tested, and the traditional nucleic acid extraction steps are usually time-consuming. Although automated nucleic acid extraction has been widely used, it still takes a long time, usually more than 30 minutes , and the steps are more complicated, and the reagent cost is relatively high. It is a simpler method to quickly release the ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6806
CPCC12Q1/6806C12Q2527/125C12Q2523/308
Inventor 李明黄海潮申保晨彭涛孙晓亮宋东亮
Owner YEASEN BIOTECHNOLOGY (SHANGHAI) CO LTD
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