Rapid nucleic acid extraction method for clinical sample detection

Abstract

The invention relates to the technical field of clinical sample detection, in particular to a rapid nucleic acid extraction method for clinical sample detection, which comprises the following steps: S1, taking 0.5-1ml of clinical biological sample, adding the clinical biological sample into a test tube filled with mixed magnetic beads, uniformly mixing, standing for 12-15 minutes, placing the test tube on a magnetic rack, standing for 3-5 minutes, and taking out the test tube; feeding into a centrifugal machine for centrifugal treatment, centrifuging for 3.5 minutes under the condition of 12000 rmp, and absorbing and discarding supernate; s2, adding 1.5 ml of normal saline into a test tube, oscillating in a vortex oscillator to scatter precipitate, sending into a centrifugal machine to carry out centrifugal treatment, centrifuging for 6-9 minutes under the condition of 12000 rmp, sucking and discarding supernate, adding 85-90 microliters of first treatment liquid into the test tube by using a pipette, adding 15-20 microliters of potassium acetate buffer solution after the solution in the test tube becomes clear, and continuously stirring for 20-30 minutes; the method can effectively solve the problems that the existing nucleic acid extraction method needs repeated centrifugation, the extraction steps are complicated, the consumed time is long, specific equipment is needed, and rapid extraction cannot be realized.

Description

technical field [0001] The invention relates to the technical field of clinical sample detection, in particular to a rapid nucleic acid extraction method for clinical sample detection. Background technique [0002] Nucleic acid is the carrier of genetic information and the material basis of gene expression. Whether it is to study the structure or function of nucleic acid, it is first necessary to quickly and effectively separate and extract the required genomic nucleic acid from complex and diverse biological samples, and the extracted Nucleic acid quality and its integrity will directly affect the subsequent experimental results. [0003] The main challenge for molecular diagnosis using nucleic acid amplification technology is the pretreatment of nucleic acid-containing materials before amplification and the complicated nucleic acid extraction steps. Currently, the widely used silica-based and magnetic bead-based technologies have many shortcomings, including efficiency To...

AI Technical Summary

Problems solved by technology

[0003] The main challenge for molecular diagnosis using nucleic acid amplification technology is the pretreatment of nucleic acid-containing materials before amplification and the complicated nucleic acid extraction steps. Currently, the widely used silica-based and magnetic bead-based technologies have many shortcomings, including efficiency Too low (≤10%); there is also the need to optimize for specific target types, resulting in the extraction process of one sample type is usually incompatible with other sample types; while the method using phenol chloroform has high recovery, but due to the The high toxicity of the components limits its applicability; the detergent-based method has a simple dissolution process, but requires high-temperature denaturation (95°C) treatment, and because the reverse transcriptase is not thermostable, it is not suitable for a single step RT-PCR

[0004] The current method of extracting nucleic acid requires repeated centrifugation, the extraction steps are complicated, time-consuming, and require specific equipment, which cannot be extracted quickly