Detection kit for novel coronavirus COVID-19 infection

Abstract

The invention discloses a detection kit for novel coronavirus COVID-19 infection and belongs to the field of in-vitro diagnosis reagents. The kit disclosed by the invention comprises primers and probes for detecting a specific gene ORF1ab of novel coronavirus COVID-19, primers and probes for detecting a positive internal reference gene GAPDH and primers and probes for detecting an exogenous monitoring gene EGFP, and sequences are separately shown in SEQ ID No. 1-9. According to the kit, on the premise that the accuracy and stability of a detection result are guaranteed, the flux of detection is increased. According to the kit, a reagent Trizol is used as a sample preservation solution, safety of living beings, completeness of nucleic acid and load capacity of virus are guaranteed. According to the kit, RNA of the exogenous monitoring gene is introduced during RNA extraction, RNA loss conditions of a whole extraction and detection process can be monitored, the normativeness of a whole operating process is guaranteed, and false negative caused by defects of any step of the operating process is avoided.

Description

technical field [0001] The invention belongs to the field of in vitro diagnostic reagents, in particular to a novel coronavirus COVID-19 infection detection kit. Background technique [0002] Since the outbreak of the novel coronavirus pneumonia, nucleic acid testing has been the "gold standard" for clinical diagnosis, recovery and discharge, and release of isolation. The currently commonly used fluorescent quantitative PCR method includes: sample collection, nucleic acid extraction, and fluorescent quantitative PCR detection. By detecting whether the submitted sample contains the specific gene of the new coronavirus, and then judging whether the sample contains the new coronavirus. [0003] However, at present, there are generally problems of false negatives and false positives in nucleic acid detection, that is, the problems of accuracy and sensitivity. This is mainly because the new coronavirus is an RNA virus, and RNase has certain stability and has a wide range of sour...

AI Technical Summary

Problems solved by technology

[0005] Regarding sample extraction, the existing methods are commonly used for direct column extraction and magnetic bead extraction. However, during the extraction process, sample loss may occur due to operational procedures or human factors, resulting in sample extraction failure. How to define sampling failure or sample extraction? False negatives caused by extraction failures, and how to define the difference in extraction efficiency between different samples need to be resolved

[0006] Regarding RT-QPCR detection, for the determination of the detection results, only for the negative detection results of virus-specific genes, it is impossible to determine whether it is a false negative caused by the failure of sampling transportation or nucleic acid extraction failure, and a positive internal standard gene needs to be set

At the same time, compared with RT and QPCR step-by-step method and dye method detection, one-step probe method detection is a more accurate and efficient detection method, but since this method does not have a step to eliminate genome contamination, the interference of genome detection caused by positive internal standard genes , can also mislead the determination of sample eligibility

Increasing the detection of internal standard genes improves the interpretation of the accuracy of the results, but also limits the detection throughput and increases the detection cost

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