A kit for extracting pathogen nucleic acids from a phlegm sample in a high-flux and automatic manner

Abstract

A kit for extracting pathogen nucleic acids from a phlegm sample in a high-flux and automatic manner is disclosed. The kit includes a phlegm digestion agent, a lysis liquid, magnetic beads, a washing liquid and an eluant. The phlegm digestion agent is a NaOH solution having a concentration of 1-6 M. The lysis liquid is an acidic solution the pH of which is not more than 3, and which is prepared from a guanidine salt having a concentration of 2-8 M, a surfactant, a pH buffer medium and a metal chelating agent. Centrifugation is not needed during extraction of nucleic acids of pathogens (including bacteria, viruses and fungi) in the phlegm sample by using the kit, thus facilitating automatic operation. The kit can extract DNAs and RNAs of bacteria, viruses and fungi in the phlegm sample so that the kit is high in universality. An extraction process is free of organic and toxic solvents so that the kit is safer. Extracted nucleic acids are high in purity so that value of a pathogen clinical sample in disease diagnosis is improved.

Description

technical field [0001] The invention relates to the technical field of biochemistry, in particular to a high-throughput kit for automatically extracting pathogen nucleic acid from sputum samples. Background technique [0002] The inner walls of the human trachea and bronchi are covered with a layer of mucous membrane, and the submucosa contains more mucus glands and serous glands. Under normal circumstances, goblet cells and glands secrete a small amount of mucus to cover the surface of the mucous membrane, which can protect the mucous membrane and keep the tracheal mucous membrane moist, so as to adhere to the dust particles and bacteria inhaled into the trachea and bronchi, and block them. It penetrates deep into the lung tissue. When the trachea, bronchi, and lungs are stimulated by harmful factors or inflamed by pathogenic bacteria, the mucous membrane of the respiratory tract is congested, edematous, a large number of inflammatory cells infiltrate, the blood vessels di...

AI Technical Summary

Problems solved by technology

This process requires repeated centrifugal washing, the operation is very complicated, and it is easy to lose a large amount of nucleic acid during the digestion and extraction process. At the same time, the removal rate of impurities is low, resulting in false negative detection; repeated centrifugation is not conducive to the automation of nucleic acid detection, and this method is not applicable The extraction of pathogen RNA in sputum is not universal, which directly limits the clinical diagnosis of diseases such as respiratory tract infections through sputum, and reduces the value of sputum, an important clinical sample, in disease diagnosis.

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