Rapid automatic extraction method and reagent for paraffin section tissue nucleic acid

Abstract

The invention discloses a rapid automatic extraction method and a reagent for paraffin section tissue nucleic acid. The reagent comprises a dewaxing agent, a protease solution, a lysate, a washing solution A, a washing solution B and an eluent. The method comprises the following steps: performing dewaxing rapidly by using the nontoxic dewaxing agent; after washing by ethanol, adding protease containing SDS and urea for cracking and digestion into liquid; transferring the liquid into an automatic magnetic bead extractor for extraction, performing cracking, magnetic bead adsorption, washing withthe washing solution A and the washing solution B and eluting with the eluent sequentially, and completing extraction of 32 or even 96 samples within a machine running time of about 30 minutes. According to the rapid automatic extraction method and the reagent for paraffin section tissue nucleic acid, after conventional first protease incubation is completed, the incubation dissolution system isheated to a high temperature and maintained for 3-5 minutes, a product is taken out and cooled to room temperature, protease is added to the reaction system, and tissue is completely dissolved and disappears after incubation for 10-30 minutes, obvious particles are absent, and the solution is clear.

Description

technical field [0001] The invention relates to a rapid and automatic extraction method and reagent for paraffin section tissue nucleic acid, belonging to the technical field of biomedicine. Background technique [0002] Tumor gene detection is currently a popular field of clinical molecular diagnosis. Through the separation and purification of tumor tissue DNA and RNA, nucleic acid detection is performed to understand the molecular characteristics of tumors, so as to predict and diagnose tumors and even judge patient prognosis, recurrence and survival, drug efficacy, etc.; Tissue embedded in slices is the most common sample for tumor genetic testing, because paraffin embedding is a classic method for long-term preservation of tissue proteins and nucleic acid molecules at room temperature, and almost all medical institutions use paraffin embedding to preserve tumor tissues; during paraffin embedding In the method, the fresh tissue is fixed with an organic solvent and gradual...

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Problems solved by technology

[0003] Generally, the thickness of paraffin section tissue is required to be between 5 and 6 microns. Thin sections are conducive to the penetration of solvents such as xylene and the digestion of protease, so that the extraction is more thorough. However, in actual operation, the thickness of paraffin sections varies in different medical institutions. And different medical institutions and different section operators will have great differences in cutting; especially when performing gene detection with a large amount of nucleic acid such as breast cancer 21 gene expression quantification, the tester will require a large amount of paraffin tissue to be cut. The number of paraffin sections and slice thickness may increase significantly; at this time, the detection operator will use the method of prolonging the protease incubation time to dissolve a large amount of paraffin tissue. Generally, the protease digestion incubation time for paraffin tissue nucleic acid extraction is 15 minutes to 3 hours, but For paraffin sections with thick slices and a large number, even if the time is extended to 24 hours, it cannot be completely dissolved, and solid tissue will still remain at the bottom of the container; if the incubation time is too long, some nucleic acid substances will be degraded and broken, and the remaining tissue Existence will not only reduce the amount of nucleic acid extracted, but also bias the representativeness of the nucleic acid extracted from the dissolved and digested part of the tissue to the whole tissue, thereby affecting the measurement results

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