Method and test strip for evaluating cross-species infection risk of coronavirus and application of test strip

Abstract

The invention belongs to the technical field of biology, and discloses a method and a test strip for evaluating cross-species infection risk of coronavirus and application of the test strip. The method comprises the following steps: capturing complete or partial spike protein of a coronavirus to be detected by taking marked complete or partial ACE2 protein of a certain vertebrate species as a probe; capturing the probes which are not combined in the step (1) by using a fixed known coronavirus spike protein S1 structural domain; detecting whether a probe which is not combined can be captured bya known coronavirus spike protein S1 structural domain or not to judge whether spike protein of the coronavirus to be detected can be combined with complete or partial ACE2 protein of a vertebrate species marked in the step (1) or not, and then evaluating the risk that the vertebrate species are infected by the coronavirus to be detected through an ACE2 receptor. The detection object is not limited to a certain specified coronavirus, and can be detected as long as the detection object can be effectively combined with the ACE2 protein.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for assessing the risk of cross-species infection of a coronavirus, a test strip and an application thereof. Background technique [0002] Severe Acute Respiratory Syndrome (SARS), Middle East Respiratory Syndrome (MARS) and Novel Coronavirus Pneumonia (COVID-19) broke out successively in 2002, 2008 and 2019. The pathogens that cause these three severe respiratory infectious diseases—SARS-CoV, MERS-CoV, and SARS-CoV-2 all belong to coronaviruses. In addition to MERS-CoV infecting humans by binding to human dipeptidyl peptidases (DPPs), SARS-CoV, SARS-CoV-2 and another coronavirus HCoV-NL63, all through the S1 domain of their surface spike protein Binding to its natural functional receptor, human angiotensin-converting enzyme 2 (ACE2), triggers infection of respiratory symptoms. Therefore, it has been proved to be an effective shortcut for the coronavirus to inf...

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Problems solved by technology

[0006] 1. Based on ethics, it is forbidden to carry out human experiments on unknown coronaviruses, and human bodies cannot be used for cross-species infection experiments;

[0007] 2. Limited to the safety level of biological laboratories, cell infection experiments or animal body transfection experiments can only be carried out in a very small number of qualified institutions, and cannot be popularized;

[0008] 3. Nucleic acid detection technology, combined with bioinformatics and structural biology, can only calculate and predict the potential hazards of unknown coronaviruses indirectly, lacking experimental evidence;

[0009] 4. The immunoassay scheme based on the specific binding of antibody antigens, the core of which is to specifically design antibodies against known coronavirus proteins, which cannot be used to assess the infection risk of unknown coronaviruses;

[0010] 5. Taking humans as an example, the purified unknown animal coronavirus S protein and human ACE2 protein were combined by SPR (surface plasmon resonance), BLI (biofilm layer surface interference technology) or ITC (isothermal calorimetric titration) for binding kinetics With (or) thermodynamic determination, the interaction between the two proteins can be accurately quantified, thereby assessing the risk of cross-species transmission, but each time an unknown coronavirus S protein needs to be exogenously expressed and multi-step purified, which takes a long period and high cost. complicated steps

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