Micro-fluidic chip for extracting total nucleic acid of bacteria in human whole blood

Abstract

The invention discloses a micro-fluidic chip for extracting total nucleic acid of bacteria in human whole blood. The chip comprises a positioning hole and a main body. The main body comprises a sampling hole formed in a sampling cavity, the sampling cavity, a one-way valve, a nucleic acid extraction material, a nucleic acid extraction cavity, a PCR reaction cavity, a buffer solution I cavity, a buffer solution II cavity, an RNA enzyme inhibitor cavity, an MIX1 cavity, a DNase I cavity, a MIX2 cavity, a 70% ethanol cavity, a buffer solution II cavity, an MIX3 cavity, a PCR Master Mix cavity, aPCR primer cavity, a PCR reaction probe cavity and a buffer solution V cavity. The chip provided by the invention can split the bacteria quickly to release RNA, and is simple and convenient to operate, so that the extraction efficiency is improved greatly; the detection limit is reduced to the lowest, so that the probability of leaked detection is avoided; the operating process of pathogenic bacteria detection is simplified, and the detection error caused by manual operations is reduced.

Description

technical field [0001] The invention relates to the technical field of molecular biomedicine, in particular to a microfluidic chip for extracting bacterial total nucleic acid from human whole blood. Background technique [0002] Bacterial infection is one of the most common diseases in the world, which can easily lead to a variety of diseases. The main clinical manifestations are: sudden chills, high fever, tachycardia, shortness of breath, rash, hepatosplenomegaly and mental and mental changes And a series of serious clinical symptoms, which are very harmful and can easily lead to death. Therefore, rapid determination of bacterial types in human whole blood has great academic research and clinical application value for understanding the diversity of bacteria in whole blood, guiding the selection and use of antibiotics, and killing toxic bacteria. In order to obtain high-quality total nucleic acids from different bacteria, pre-sample processing, extraction methods and selec...

AI Technical Summary

Problems solved by technology

In addition, it must be pointed out that for different bacteria, the results presented by the same method often vary greatly

There are many published methods for extracting bacterial total nucleic acid, but most methods not only need to use some toxic reagents, but also are time-consuming, complicated, expensive, and the extraction effect is not good.

In addition, many bacteria are difficult to be destroyed and lysed because they are protected by cell walls. For example, Gram-negative bacteria have a relatively single cell wall composition, a fragile structure, and are easily broken; while Gram-positive bacteria have thicker cell walls. The chemical structure is conservative and fixed, the three-dimensional cell wall structure is tightly cross-linked, the cells are difficult to break, and the extraction is difficult

Moreover, the existing methods cannot detect the number of bacteria with extremely low content, resulting in the possibility of missed detection of pathogenic bacteria

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