Rapid PCR amplification and CRISPR visual detection device and method

Abstract

The invention relates to a rapid PCR amplification and CRISPR visual detection device and method. The visual detection device comprises a closed module and a heating module, wherein the closed modulecomprises a lower carrier, an upper cover body, a plurality of PCR amplification chambers arranged in the lower carrier and a plurality of CRISPR reaction chambers of the corresponding number; the amplification chambers are communicated with the corresponding CRISPR reaction chambers through connecting channels; the upper cover body is connected with the lower carrier in a sealed manner and all the chambers and the connecting channels are enabled to be sealed; and the heating module is used for heating the PCR amplification chambers and / or the CRISPR reaction chambers. Compared with the priorart, the device has the advantages that the specific surface area is large, and the heat transfer efficiency is improved, so that the PCR amplification time is greatly shortened, and the total detection time is further shortened to be within a few minutes. According to the invention, amplification of a plurality of PCR systems and CRISPR visual detection can be realized, and the temperature-controllable heating module is integrated in the detection device, so that the detection is more convenient, and the detection time is shortened.

Description

technical field [0001] The invention belongs to the field of nucleic acid amplification and visual detection, and in particular relates to a rapid PCR amplification and CRISPR visual detection device and detection method. Background technique [0002] Since the birth of polymerase chain reaction (PCR) technology in 1985, nucleic acid amplification detection technology has been rapidly applied in various fields such as medical treatment, agriculture, and environmental sanitation due to its high detection sensitivity. For the detection of nucleic acid amplification products, the visual detection method has great advantages in the field and rapid detection of nucleic acids because it does not require equipment and can interpret the results with naked eyes. Existing visual detection methods usually use high-concentration fluorescent intercalating dyes (SYTO 9, SYBR GREEN I, ethidium bromide, propidium iodide, etc.) Fluorescence to achieve visual detection; or use calcein, methy...

AI Technical Summary

Problems solved by technology

Since the detection sensitivity of the CRISPR method alone is not high, it is usually combined with nucleic acid amplification

However, because the CRISPR system is not compatible with the nucleic acid amplification system or the reaction temperature is different from the nucleic acid amplification system, the cleavage efficiency of the CRISPR / Cas enzyme is greatly reduced or inactivated. Therefore, researchers usually divide nucleic acid amplification and CRISPR detection into two steps. After the nucleic acid amplification is completed, the amplification product is transferred to the CRISPR system for detection

The operation is complicated and cumbersome, and due to the large output of amplicons, opening the cover after the amplification is completed can easily cause the problem of amplicon aerosol pollution, resulting in false positives in subsequent detection results, making the detection results unreliable

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