Reagent device, reagent and method for extraction and detection of nucleic acid

Abstract

The invention discloses a reagent device, a reagent and a method for extraction and detection of nucleic acid. The reagent device includes a nucleic acid extraction chamber and a nucleic acid detection chamber, wherein a first reagent is pre-packaged in the nucleic acid extraction chamber and is used for extracting and purifying the nucleic acid, and a second reagent is pre-packaged in the nucleicacid detection chamber and is used for detecting the nucleic acid. The first reagent includes a cell lysis solution, a nucleic acid washing solution and a nucleic acid elution liquid; the second reagent includes a plurality of PCR freeze-dried reagents for detecting a plurality of pathogens. The PCR freeze-dried reagents include a multiple PCR reaction buffer solution, a DNA polymerase, specificprimers of the pathogens, and probes, and a specific primer and a probe used for internal quality control. The sealed reagent device can automatically achieve extraction, purification and detection tofifteen respiratory tract pathogens, is high in reagent sensitivity, has simple operations and is easy to promote in clinical application.

Description

technical field [0001] The invention relates to the field of nucleic acid extraction and purification, in particular to a device, reagent and method for nucleic acid extraction and detection of various pathogens in a closed system. Background technique [0002] Acute respiratory infection is one of the most common infectious diseases. In developing countries, acute respiratory disease accounts for 75% of acute morbidity, and 80% of acute upper respiratory diseases and most of lower respiratory diseases are caused by pathogens other than bacteria. Among them, respiratory viruses are the most common [1] (Mahony JB. Detection of Respiratory Viruses by Molecular Methods [J]. Clinical Microbiology Reviews, 2008, 21 (4): 716-747.). Common respiratory viruses include: Influenza A virus (FluA), Influenza B virus (FluB), Respiratory syncytial virus (Respiratory syncytial virus, RSV), Human Metapneumovirus (Human Metapneumovirus, HMPV), adenovirus (Adenovirus, AdV), human rhinovirus ...

AI Technical Summary

Problems solved by technology

The entire detection process is cumbersome and time-consuming

Moreover, the extraction and purification efficiencies of different extraction reagents vary greatly, resulting in large differences in the detection results of downstream multiplex fluorescent PCR and poor stability.

In addition, most of these detection kits require low-temperature storage and transportation, and have high requirements for the storage and transportation environment of reagents.

Not only that, the use of reagents has high requirements on the professional skills of operators and hardware equipment, which is not conducive to clinical promotion

Images