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Novel coronavirus detection kit

A detection kit, coronavirus technology, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve problems such as long operation time, false negatives, and high technical level requirements for testing personnel

Active Publication Date: 2021-08-31
SHENZHEN YHLO BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, since the development of the epidemic, the problem of missed detection of nucleic acid detection reagents has been repeatedly exposed, and the positive detection rate is only 30% to 50%, mainly due to the limitations of detection methods.
Nucleic acid detection generally collects nasopharyngeal swabs, sputum, blood, and feces of suspected patients, and extracts RNA fragments from them for fluorescent RT-PCR. During the sampling process, a large number of human cells and bacteria will be mixed in, resulting in the abundance of RNA to be tested Low, the detection system cannot detect, eventually leading to false negatives
At the same time, except for blood drawing, other sampling methods will cause the sampling personnel to be exposed to the environment that may contain pathogens, and there is a potential risk of infection
Moreover, the preparation process of nucleic acid detection samples requires many steps and long operation time, which requires high technical level of testing personnel.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0040] In a specific example, the preparation method of the novel coronavirus protein-magnetic bead complex includes the following steps: use 3mM~7mM sodium dodecyl sulfate solution to treat the novel coronavirus protein to obtain a denatured antigen, and then use the denatured antigen to coat the magnetic beads. beads. Preferably, the concentration of SDS is 4mM-6mM. Due to the large variety of antigens and the complex conformation, in the preparation process of the protein-magnetic bead complex, by using a certain concentration of SDS to treat the recombinant 2019-nCoV-related antigen, the antigenic epitope is released, which increases the number of antigens that are different from the 2019-nCoV in different samples. The possibility of virus IgG antibody binding improves the sensitivity of detection and further reduces the possibility of false negative or missed detection.

[0041] Aiming at the defects of the nucleic acid detection method and possible omissions in the chem...

Embodiment 1

[0044] 1. Antigen preparation

[0045] According to the gene sequence of the new coronavirus and the composition of its shell, the 293 expression system was used to express and purify the proteins that make up the structural proteins of the new coronavirus (spike protein Spike-S1 subunit, Spike-S2 subunit, nucleocapsid protein, capsid protein, membrane protein) as the coating antigen.

[0046] For example, the expression and purification of S1 protein:

[0047] Select the RBD (receptor binding domain) protein of the new coronavirus spike protein, the amino acid sequence is:

[0048] NITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFV IRGDEVRQIAPGQTGKIADYNYKLPDDFTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFERDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQPTFENOVGYQPYLLRVVVVTV(IDNLCPFGYQPYLLRVVVST)

[0049] Signal peptide, the amino acid sequence is: MFVFLVLLPLVSSQCV (SEQ ID NO: 2);

[0050] Transmembrane domain, the amino acid sequence is: ELWLVLVAVGAGLLLLLGLIILLL (SEQ ID NO: 3);...

Embodiment 2

[0085] This example is basically the same as Example 1, except that different concentrations of SDS (2mM, 7mM, 10mM) were used in the preparation process of the antigen-magnetic bead complex.

[0086]

[0087]

[0088] 10 positive specimens were tested using kits obtained under conditions of different concentrations of SDS. As can be seen from the above table, when 2mM SDS was used, the test results of individual samples were negative due to insufficient antigen exposure during processing. When 5mM SDS was used, the treatment effect was the best, and all 10 positive samples could be detected. However, when 7mM and 10mM are used, due to the high concentration used, the protein is completely denatured, and the antigenic site is destroyed, and some positive test results are negative.

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Abstract

The invention relates to a novel coronavirus detection kit. The novel coronavirus detection kit comprises an antigen component and a luminous component, the antigen component contains a novel coronavirus protein-magnetic bead compound, and the novel coronavirus protein-magnetic bead compound comprises an S1 protein-magnetic bead compound, an S2 protein-magnetic bead compound, an E protein-magnetic bead compound, an M protein-magnetic bead compound and an N protein-magnetic bead compound. The luminous component contains an anti-human IgG antibody, and a luminous marker is marked on the anti-human IgG antibody. The novel coronavirus detection kit disclosed by the invention is based on a two-step indirect immunoassay principle of direct chemiluminescence, the concentration of a novel coronavirus IgG antibody in a to-be-detected sample can be quickly judged by utilizing a corresponding chemiluminescence tester, and by coating all novel coronavirus antigens which possibly generate antibodies to the surfaces of magnetic beads, the missing detection caused by different antibodies generated in bodies of individual patients is avoided to the greatest extent.

Description

technical field [0001] The invention relates to the technical field of virus detection, in particular to a novel coronavirus detection kit. Background technique [0002] The new type of coronavirus (SARS-CoV-2, also known as 2019-nCoV, HCoV-19) belongs to the genus β coronavirus, the particles are round or oval in diameter 60nm ~ 140nm, the genetic characteristics are significantly different from SARS, And it has a higher infectious ability than SARS. Based on epidemiological surveys, the incubation period of COVID-19 is 1 to 14 days, mostly 3 to 7 days. The main symptoms are fever, dry cough, and fatigue. Severe patients often have dyspnea and / or hypoxemia one week after the onset In severe cases, it can rapidly progress to acute respiratory distress syndrome, septic shock, difficult-to-correct metabolic acidosis, coagulation dysfunction, and multiple organ failure. Therefore, how to quickly diagnose suspected cases and how to effectively monitor the progress of the disea...

Claims

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Application Information

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IPC IPC(8): G01N33/569G01N33/543
CPCG01N33/56983G01N33/54326
Inventor 程方明马腾腾钱纯亘徐天旺胡鹍辉
Owner SHENZHEN YHLO BIOTECH
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