Detection kit for novel coronavirus COVID-19 infection

A technology for COVID-19 and coronavirus, applied in the field of new coronavirus COVID-19 infection detection kits, can solve the problems of increased detection costs, increased internal standard gene detection, inability to determine, etc., to ensure detection sensitivity and ensure biological safety. The effect of increasing the detection sensitivity

Pending Publication Date: 2020-07-31
武汉生命之美科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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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

Method used

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  • Detection kit for novel coronavirus COVID-19 infection
  • Detection kit for novel coronavirus COVID-19 infection
  • Detection kit for novel coronavirus COVID-19 infection

Examples

Experimental program
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Effect test

Embodiment 1

[0086] The above protocol was used to collect nasal swabs and oropharyngeal swabs from 11 individuals, as well as cerebrospinal fluid and blood samples from 8 patients, RNA extraction, and fluorescent quantitative PCR detection. The results are shown in Table 1 and figure 2 .

[0087] The Ct value of the positive internal reference gene GAPDH detection of nasal swab and oropharyngeal swab is between 12-25 (Table 5), the Ct value of the positive internal reference gene GAPDH detection of cerebrospinal fluid sample is between 23-31, and the positive internal reference gene GAPDH detection of blood sample The Ct value detected by the internal reference gene GAPDH was between 16-18 (Table 6), which met the requirements for sample preservation and extraction; The new coronavirus, which is not found in the preservation and extraction of existing methods, fully proves the reliability, stability and sensitivity of Trizol reagent for preserving samples and performing nucleic acid extr...

Embodiment 3

[0095] Using the above fluorescent quantitative PCR detection method to carry out single gene detection and double PCR detection on GAPDH, ORF1ab standard products, the results are shown in Table 7 and image 3. It can be seen from the results that the Ct value of the single-gene detection and the Ct value of the double PCR detection are almost the same for the standards with the same number of molecules, indicating that the specificity of the primers is better, and there is no significant difference between the two groups of primers and probes. Influence, the linear relationship is also very good.

[0096] Table 7

[0097]

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PUM

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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...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/6806C12Q1/6851C12Q1/6848C12R1/93
CPCC12Q1/701C12Q1/6806C12Q1/6851C12Q1/6848C12Q2600/166C12Q2600/16C12Q2527/125C12Q2531/113C12Q2545/113C12Q2537/143C12Q2521/531
Inventor 张翼孙月张倩陈栋程超刘圆圆王启
Owner 武汉生命之美科技有限公司
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