A method and system for detecting viral or pseudoviral infection of cells.

Abstract

This invention discloses a method for detecting viral or pseudoviral cell infection, comprising: applying a sinusoidal voltage to an attached membrane in patch-clamp mode and detecting the corresponding membrane current; inducing viral or pseudoviral cell infection by binding of viruses or pseudoviruses in the electrode fluid to receptors on the attached membrane; performing phase discrimination demodulation on the corresponding membrane current signal using the phase of the input sinusoidal voltage as a standard to obtain the imaginary and real parts of the membrane current and calibrating and standardizing them in real time; calculating the membrane capacitance of the attached membrane based on the real and imaginary parts of the current signal, based on the calibration and standardization; identifying viral or pseudoviral cell infection events and their modes based on the dynamic characteristics and polarity of the membrane capacitance changes; and estimating the size of the virus or pseudovirus based on the magnitude of the membrane capacitance changes. This invention enables real-time, high-precision detection of various viruses or pseudoviruses infecting host cells via envelope-cell membrane fusion or endocytosis in acutely isolated cells, cultured cells, and isolated tissue cells.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of biological and medical detection, in particular to a virus or pseudo-virus infection cell detection method and system. BACKGROUND

[0002] The research of viruses and the development of their therapeutic drugs are all started from the aspects of virus infection, reverse transcription, replication and self-packaging. Taking the new coronavirus as an example, the detection and research of the first line of defense, i.e. the new coronavirus infection of cells, is crucial to the prevention and control of the new coronavirus epidemic. The diameter of a virus is about tens to hundreds of nanometers, and the process of infection of cells is completed within milliseconds. The current technical means for virus detection and observation of its infection of cells in the field include electron microscopy observation, high-resolution fluorescence microscopic imaging based on protein expression, immunoblotting and nucleic acid detection methods.

[0003] Among them, the electron microscopy observation method has high spatial accuracy and intuitiveness, and can observe the infection event of a single virus at a low probability, but this method cannot realize the observation of living cells, and can only complete the static detection of the morphology of cells or subcells at a certain time point. There are limitations such as long and tedious processing process, easy contamination and damage of samples. High-resolution fluorescence microscopic imaging based on protein expression can detect the motion trajectory of a single virus particle in real time at a low time resolution (sub-second level), but the observation sample is limited to cultured cells, and the virus must be genetically or proteinically modified. Immunoblotting and nucleic acid detection methods have limited signal accuracy and no time and space information, making it difficult to accurately and real-time monitor the dynamic process of virus infection of living cells.

[0004] In summary, the existing methods are difficult to accurately and real-time monitor the dynamic process of virus infection of living cells due to the limitations of the technology itself, or may change the properties of the virus due to the need for genetic or protein modification of the virus. In addition, the above methods cannot directly observe the virus infection on human living tissues, so there is still a distance between the corresponding observation and the real human pathological phenomena, which limits the diagnosis and treatment of virus-related diseases and the drug development process. The above technical problems need to be solved by developing an accurate and real-time method for detecting virus infection of living cells from a new perspective and with innovative means. SUMMARY

[0005] In view of the problems existing in the prior art virus infection cell detection technology, the present application realizes real-time and high-precision detection of various viruses or pseudo-viruses with replaced RNA or DNA on the living cells of acute isolated cells, cultured cells and isolated tissues of animals and humans.

[0006] The technical solution of the present application is as follows:

[0007] According to one aspect of the present application, there is provided a method for detecting virus or pseudo-virus infected cells.

[0008] The method for detecting virus or pseudo-virus infected cells comprises:

[0009] The patch-clamp mode is established to enable the virus or pseudo-virus in the electrode internal solution to bind with the receptors on the membrane patch contained in the electrode to induce virus or pseudo-virus infected cells;

[0010] A sinusoidal voltage is applied to the attached membrane patch of the cells under the patch-clamp mode, and the corresponding membrane current is detected;

[0011] The phase of the input sinusoidal voltage is used as a standard to demodulate the corresponding membrane current signal, and the imaginary part signal and the real part signal of the membrane current are obtained, and real-time calibration and scaling are performed;

[0012] The membrane capacitance change caused by the virus or pseudo-virus infected cells is detected and calculated based on the real part signal and the imaginary part signal of the current under the calibration and scaling;

[0013] The virus or pseudo-virus infected cell event and the mode thereof are identified according to the kinetic characteristics and polarity of the membrane patch capacitance change, and the size of the virus or pseudo-virus is estimated according to the size of the membrane patch capacitance change.

[0014] In addition, the method for detecting virus or pseudo-virus infected cells further comprises: on the basis of realizing the cell attached patch-clamp recording of the electrode internal and external giant resistance electrical isolation, configuring a phase demodulation module and using the signal generation function thereof to apply a high-frequency sinusoidal voltage to the attached membrane patch and detect the corresponding membrane current.

[0015] The phase of the input sinusoidal voltage is used as a standard to demodulate the corresponding membrane current signal, and the imaginary part signal and the real part signal of the membrane current are obtained, and real-time calibration and scaling are performed;

[0016] The membrane capacitance change caused by the virus or pseudo-virus infected cells is detected and calculated based on the real part signal and the imaginary part signal of the current under the calibration and scaling;

[0017] According to the current real part and imaginary part signals, the changes of the imaginary part signal and the real part signal are directly related to the capacitance change through calibration, and the scale coefficient K is determined through scaling, and the membrane capacitance change is calculated by the following calculation formula:

[0018]

[0019] In the formula, Cv is the membrane capacitance change of the detection area, Re is the change of the current real part signal, and Im is the change of the current imaginary part signal.

[0020] According to the kinetics and polarity of the membrane patch capacitance change, the virus or pseudo-virus infection cell event and its mode are identified, wherein if the membrane capacitance step rises, the infection cell event in the fusion mode of the virus or pseudo-virus envelope and the cell membrane is identified; if the membrane capacitance step falls, the infection cell event in the endocytosis mode of the virus or pseudo-virus particle is identified. The size of the virus or pseudo-virus is estimated according to the size of the membrane patch capacitance change, and the size of the virus or pseudo-virus (envelope area or endocytosis envelope) is estimated according to the ratio of the size of the membrane patch capacitance change to the capacitance per unit membrane area.

[0021] According to another aspect of the present application, a virus or pseudo-virus infection cell detection system is provided.

[0022] The virus or pseudo-virus infection cell detection system comprises:

[0023] An infection induction unit is configured to establish a patch clamp mode to enable the virus or pseudo-virus in the electrode internal liquid to combine with the receptors on the membrane patch contained in the electrode to induce the virus or pseudo-virus to infect the cell.

[0024] A membrane current detection unit is configured to apply a sinusoidal voltage to the attached membrane patch of the cell in the patch clamp mode and detect the corresponding membrane current.

[0025] A phase demodulation unit is configured to demodulate the corresponding membrane current signal in phase with the phase of the input sinusoidal voltage to obtain the imaginary part signal and the real part signal of the membrane current, and to calibrate and scale in real time.

[0026] A membrane capacitance calculation unit is configured to detect and calculate the membrane capacitance change caused by the virus or pseudo-virus infection cell on the basis of the calibrated and scaled current real part signal and current imaginary part signal.

[0027] An event identification unit is configured to identify the virus or pseudo-virus infection cell event and its mode according to the kinetics and polarity of the membrane patch capacitance change, and to estimate the size of the virus or pseudo-virus according to the size of the membrane patch capacitance change.

[0028] The membrane current detection unit is configured to apply a high-frequency sinusoidal voltage to the attached membrane patch and detect the corresponding membrane current by using the signal generation function of the phase demodulation module on the basis of the cell attached patch clamp recording in which the internal and external electrodes are electrically isolated by a large resistance.

[0029] The phase demodulation unit is configured to detect the corresponding membrane current after the voltage is applied to the cell membrane patch in the patch clamp mode, and to demodulate the membrane current to obtain the current real part signal and the current imaginary part signal of the membrane current when the corresponding membrane current signal is demodulated in phase with the phase of the input sinusoidal voltage.

[0030] Wherein, the membrane capacitance calculation unit, when detecting and calculating the membrane capacitance change caused by the virus or pseudo-virus invading the cells according to the current real part signal and the current imaginary part signal on the basis of calibration and scaling, according to the current real part and imaginary part signals, makes the change of the imaginary part signal and the real part signal directly related to the capacitance change through calibration, determines the scale coefficient K through scaling, and calculates the membrane capacitance change by the following calculation formula:

[0031]

[0032] In the formula, Cv is the cell membrane capacitance change of the detection area, Re is the change of the current real part signal, and Im is the change of the current imaginary part signal.

[0033] Wherein, the event recognition unit, when recognizing the virus or pseudo-virus invading cell events and the way according to the dynamic characteristics and polarity of the membrane capacitance change, if the membrane capacitance step rises, it is recognized as the virus or pseudo-virus envelope fusion way of invading cell events; if the membrane capacitance step falls, it is recognized as the virus particle or pseudo-virus particle endocytosis way of invading cell events. When estimating the size of the virus or pseudo-virus according to the size of the membrane capacitance change, the size of the virus or pseudo-virus (capsid membrane area or endocytosis envelope) is estimated according to the ratio of the size of the membrane capacitance change to the unit membrane area capacitance.

[0034] Beneficial effects:

[0035] The present application uses an electrical method to realize real-time monitoring of the dynamic process of wild and various mutant viruses or pseudo-viruses invading cells under physiological conditions and pharmacological intervention conditions, and the precision reaches the observation of a single virus or pseudo-virus invading cell event; the detection technology can also be used for high-throughput large-scale rapid drug screening system by expanding the measurement channel and capacity, and the detection technology of the present application can also be used for direct and high-precision observation of viruses or pseudo-viruses invading artificial membrane systems, cultured cell systems and human tissue living cells, and has great potential in virus prevention and treatment.

[0036] Compared with the electron microscope imaging method, the observation of living cells cannot be realized, only the static detection of cell or subcellular morphology at a certain time point can be completed, and the process is long and tedious, the sample is easy to be contaminated and damaged, and the like. Compared with the high-resolution fluorescence microscopic imaging based on protein expression, the observation sample is limited to cultured cells, and the virus must be genetically or proteinically modified. Compared with the immunoblotting and nucleic acid detection methods, there is no time and space information, and the signal precision is limited, and it is difficult to accurately and real-timely monitor the dynamic process of virus infection of living cells. The present application realizes the dynamic, direct and high-precision real-time detection of the dynamic process of virus infection of living cells on the premise of no sample contamination and damage and no genetic or protein modification of the virus on acute isolated cells, cultured cells and isolated tissue living cells, especially isolated human tissue living cells. It is the closest observation method of virus infection of living cells to the real human pathological condition. BRIEF DESCRIPTION OF DRAWINGS

[0037] In order to more clearly illustrate the technical solutions in the embodiments of the present application or the prior art, the drawings needed in the embodiments will be briefly introduced below. Obviously, the drawings in the following description are only some embodiments of the present application, and other drawings can be obtained by those skilled in the art without creative labor on the basis of these drawings.

[0038] Figure 1 It is a detection method flowchart of the false new coronavirus infection of cells according to the embodiments of the present application;

[0039] Figure 2 It is a structure diagram of the detection system of the false new coronavirus infection of cells according to the embodiments of the present application;

[0040] Figure 3 It is a flowchart of single false new coronavirus infection event detection in ACE2 / TMPRSS2-expressed cell lines according to the embodiments of the present application;

[0041] Figure 4 It is a flowchart of single false new coronavirus infection event detection in HEK293T, Coca-2 and other cell lines transfected with ACE2 and TMPRSS2 according to the embodiments of the present application;

[0042] Figure 5 It is a flowchart of single false new coronavirus infection event detection of alveolar cells in human lung tissue according to the embodiments of the present application. DETAILED DESCRIPTION

[0043] With reference to the drawings and the embodiments of the present application, the technical solutions in the embodiments of the present application will be described clearly and completely. Obviously, the described embodiments are only a part of the embodiments of the present application, rather than all the embodiments of the present application. Based on the embodiments of the present application, all other embodiments obtained by a person of ordinary skill in the art belong to the scope of protection of the present application.

[0044] According to an embodiment of the present application, a method and system for detecting false new coronavirus infection of HEK293T cells expressing ACE2 and TMPRSS2 proteins are provided.

[0045] As shown in the method for detecting false new coronavirus infection of cells according to an embodiment of the present application, the method comprises: Figure 1

[0046] Step S101, establish an attached patch clamp mode so that the false new coronavirus in the electrode internal solution can bind to the receptors on the patch contained in the electrode to induce false new coronavirus infection of cells;

[0047] Step S103, apply an input sinusoidal voltage to the attached patch locally under the attached patch clamp mode and detect the corresponding membrane current;

[0048] Step S105, phase demodulate the corresponding membrane current signal with the phase of the input sinusoidal voltage as the standard, obtain the imaginary part and real part of the membrane current, and calibrate and scale in real time;

[0049] Step S107, detect and calculate the membrane capacitance change caused by the false new coronavirus infection of cells based on the current real part signal and the current imaginary part signal on the basis of calibration and scaling;

[0050] Step S109, identify the false new coronavirus infection of cells event and its mode according to the kinetic characteristics and polarity of the patch capacitance change, and estimate the size of the false new coronavirus according to the size of the patch capacitance change.

[0051] In one embodiment, the method for detecting false new coronavirus infection of cells further comprises: applying a sinusoidal voltage to the patch contained in the electrode by configuring a phase demodulation module using the signal generating function of the phase-locked amplifier; recording the membrane current induced by applying a voltage to the patch under the attached patch clamp mode, and phase demodulating the corresponding membrane current signal with the phase of the input sinusoidal voltage as the standard using the phase demodulation function of the phase-locked amplifier to obtain the imaginary part and real part of the membrane current;

[0052] In one embodiment, according to the membrane current real part and imaginary part signals, the changes of the imaginary part and real part signals are directly related to the capacitance change by calibration, and the scale coefficient K is determined by scaling, and the membrane capacitance change is calculated by the following calculation formula:

[0053] ​

[0054] Where Cv represents the change in cell membrane capacitance in the detection region, Re represents the change in the real part of the current signal, and Im represents the change in the imaginary part of the current signal.

[0055] In one embodiment, the virus or pseudovirus infection event identification and analysis module identifies pseudo-SARS-CoV-2 cell infection events and their modes based on the dynamic characteristics and polarity of membrane capacitance changes. It estimates the scale of the pseudo-SARS-CoV-2 based on the magnitude of the membrane capacitance change, including: if the membrane capacitance increases stepwise, it is identified as an infection event involving the fusion of the pseudo-SARS-CoV-2 envelope with the cell membrane; if the membrane capacitance decreases stepwise, it is identified as an infection event involving the endocytosis of pseudo-SARS-CoV-2 particles. The scale of the pseudo-SARS-CoV-2 (enveloped area or endocytic envelope) is estimated based on the ratio of the magnitude of the membrane capacitance change to the capacitance per unit membrane area.

[0056] To facilitate understanding of the above technical solutions of the present invention, the following detailed description and experiments will be conducted to illustrate the above technical solutions of the present invention.

[0057] In one example of detecting pseudo-SARS-CoV-2 infection in cultured cells expressing the SARS-CoV-2 receptor protein ACE2 and the accessory protein TMPRSS2, the experimental procedure for detecting pseudo-SARS-CoV-2 infection in these cultured cells is as follows: Figure 3 As shown: HEK293T cells expressing the SARS-CoV-2 receptor protein ACE2 and the accessory protein TMPRSS2 were used as candidate host cells. During the experiment, the candidate host cells (populations) were transferred into the recording chamber containing extracellular fluid in the patch-clamp electrophysiology platform.

[0058] The extracellular fluid formulation is as follows:

[0059] Ingredients g / L NaCl 9 KCl 0.4 MgCl2 0.5 CaCl2 0.3 HEPES 2.5 D-Glucose 2 Distilled water to 1000 ml, adjust pH to 7.4

[0060] The detection was performed in a cell-attached patch-clamp mode: a glass microelectrode containing a test solution of spurious SARS-CoV-2, with a tip diameter of approximately 1 μm, was clamped to a conventional electrophysiological amplifier probe. A negative pressure airflow was applied to ensure a tight seal between the tip and the cell membrane surface, achieving giant resistance electrical isolation between the inside and outside of the electrode to significantly reduce background noise and achieve a high signal-to-noise ratio measurement. The detection system for spurious SARS-CoV-2 cell infection is as follows: Figure 2 As shown, it consists of an electrophysiological signal acquisition module, a phase detection and demodulation module, a real-time phase calibration module, a capacitance signal calibration module, a membrane capacitance detection module, and a virus or pseudovirus infection of cell event recognition and analysis module. Specifically, it includes:

[0061] Electrophysiological signal acquisition module: Based on a patch-clamp amplifier, it realizes cell-attached patch-clamp recording with giant resistance electrical isolation inside and outside the electrode. It acquires the attached patch current in a high-frequency sinusoidal voltage clamp mode modulated by DC level, amplifies it with high gain, and outputs it to the phase detection demodulation module.

[0062] Phase detection and demodulation module: The high-frequency sine wave (non-RF sine wave above 5kHz) voltage is output as a modulation signal to the electrophysiological signal acquisition module, and this is used as a phase-locked standard to perform phase detection and demodulation on the current signal input from the electrophysiological signal acquisition module to obtain the imaginary and real parts of the signal;

[0063] Phase real-time calibration module: used to realize real-time compensation of system distributed conductance and capacitance, so that the changes in the imaginary and real parts of the signal are directly related to the changes in membrane capacitance, while maintaining the optimal signal-to-noise ratio;

[0064] Membrane capacitance signal calibration module: Used to calibrate the detected membrane capacitance signal. The scaling coefficient K is determined through calibration.

[0065] Membrane capacitance detection module: The imaginary and real parts of the attached membrane current signal output by the phase detection and demodulation module are converted from analog to digital using a high-speed analog-to-digital converter. The attached membrane capacitance and conductance signals are detected by combining software and hardware based on real-time phase calibration and signal scaling, and the corresponding information is stored in the computer.

[0066] The module for identifying and analyzing viral or pseudovirus cell infection events uses software to post-process the attached membrane capacitance and conductance signals stored in the computer. When identifying pseudovirus cell infection events and their modes based on the kinetic characteristics and polarity of membrane capacitance changes, a step increase in membrane capacitance indicates infection via fusion of the pseudovirus envelope and cell membrane; a step decrease indicates infection via endocytosis. When estimating the size of the pseudovirus based on the magnitude of membrane capacitance changes, the size of the pseudovirus (enveloped area or endocytic envelope) is estimated by the ratio of the change in membrane capacitance to the capacitance per unit membrane area. This allows for analysis of the size, infection mode, kinetic process, and temporal characteristics of the pseudovirus.

[0067] When applied to different scenarios, taking the detection system described in this invention as an example, the experimental procedure for detecting the invasion of cells by the fake COVID-19 virus is as follows:

[0068] 1) Artificial cell membrane

[0069] like Figure 3 As shown, liposomes embedded with ACE2 / TMPRSS2 membrane proteins were used to measure and detect pseudo-SARS-CoV-2 cell infection events using cell-attached membrane capacitance measurement technology.

[0070] 2) Cultured cell lines

[0071] like Figure 4As shown, ACE2 and TMPRSS2 were transfected into HEK293T and Coca-2 cell lines, and cell adhesion-based lower membrane capacitance measurement was used to detect pseudo-SARS-CoV-2 cell invasion events.

[0072] Preparation of live human lung tissue sections for the detection of spurious SARS-CoV-2 infection.

[0073] like Figure 5 As shown, the live pathological tissue removed from the patient's lungs during surgery was used as a sample. The sample was prepared into live lung tissue sections with a thickness of 150-400 micrometers using a vibrating microtome. Cell adhesion membrane capacitance was then performed on alveolar epithelial cells to detect single pseudo-SARS-CoV-2 cell invasion events using membrane capacitance measurement technology.

[0074] Therefore, by means of the above technical solution, the present invention uses electrical methods to achieve real-time detection of the infection of host cells by pseudo-SARS-CoV-2 in the manner of envelope-cell membrane fusion and pseudo-SARS-CoV-2 particle endocytosis at the levels of artificial cell membranes, isolated cells of animals and humans, cultured cells and living tissues, with the accuracy reaching the observation of a single pseudo-SARS-CoV-2 cell infection event.

[0075] The detection technology of this invention can also be used for direct, high-precision observation of other viruses or pseudoviruses infecting artificial membrane systems, cultured cell systems, and living human tissue cells. This detection technology can also be used for high-throughput, large-scale, and rapid drug screening by expanding measurement channels and capacity, and has great potential in virus prevention and control.

[0076] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A method for detecting infection of a cell by a virus or a pseudovirus, characterized in that, The method comprises the following steps: A patch-clamp mode is established so that viruses or pseudo-viruses in the liquid in the electrode can bind to receptors on the membrane patch contained in the electrode to induce virus or pseudo-virus to infect cells; A sinusoidal voltage is applied to the local patch of the cell in the patch-clamp mode, and the corresponding membrane current is detected; The phase demodulation of the corresponding membrane current signal is performed based on the phase of the input sinusoidal voltage, and the imaginary part signal and the real part signal of the membrane current are obtained, and real-time calibration and scaling are performed; The membrane capacitance change caused by the virus or pseudo-virus infecting the cells is detected and calculated based on the current real part signal and the current imaginary part signal based on the calibration and scaling; The virus or pseudo-virus infection event and its mode are identified according to the dynamic characteristics and polarity of the membrane capacitance change, and the size of the virus or pseudo-virus is estimated according to the size of the membrane capacitance change.

2. The method for detecting viral or pseudoviral infection of cells according to claim 1, characterized in that, The method further comprises the following steps: On the basis of realizing the cell patch-clamp recording of the electrode inside and outside to achieve giant resistance electrical isolation, a phase demodulation module is configured, and the signal generation function of the phase demodulation module is used to apply a high-frequency sinusoidal wave voltage to the patch and detect the corresponding membrane current.

3. The method for detecting viral or pseudoviral infection of cells according to claim 1, characterized in that, The phase demodulation of the corresponding membrane current signal is performed based on the phase of the input sinusoidal voltage, and the imaginary part and the real part of the membrane current are obtained.

4. The method for detecting viral or pseudoviral infection of cells according to claim 1, characterized in that, The method comprises the following steps: The membrane current corresponding to the voltage applied to the cell patch in the patch-clamp mode is detected, and the phase demodulation of the membrane current is performed to obtain the current real part signal and the current imaginary part signal of the membrane current. ； The membrane capacitance change caused by the virus or pseudo-virus infecting the cells is detected and calculated based on the current real part signal and the current imaginary part signal based on the calibration and scaling, which comprises:

5. The method of claim 1, wherein the virus or pseudo-virus is selected from the group consisting of: adenovirus, herpes simplex virus, influenza virus, human immunodeficiency virus, and pseudovirus. According to the current real part and imaginary part signals, the change of the imaginary part signal and the real part signal is directly related to the capacitance change through calibration, and the scale coefficient K is determined through scaling, and the membrane capacitance change is calculated by the following calculation formula: Wherein, Cv is the cell membrane capacitance change of the detection area, Re is the change of the current real part signal, and Im is the change of the current imaginary part signal. The virus or pseudo-virus infection event and its mode are identified according to the dynamic characteristics and polarity of the membrane capacitance change, and the size of the virus or pseudo-virus is estimated according to the size of the membrane capacitance change, which comprises: If the membrane capacitance step rises, it is identified as a virus or pseudo-virus membrane fusion mode of infecting cells; 6. A system for detecting infection of a cell by a virus or a pseudovirus, characterized in that, If the membrane capacitance step drops, it is identified as a virus or pseudo-virus endocytosis mode of infecting cells; The size of the virus or pseudo-virus is estimated according to the ratio of the size of the membrane capacitance change to the capacitance per unit membrane area. The method comprises the following steps: An infection induction unit is configured to establish a patch-clamp mode so that viruses or pseudo-viruses in the liquid in the electrode can bind to receptors on the membrane patch contained in the electrode to induce virus or pseudo-virus to infect cells; A membrane current detection unit is configured to apply a sinusoidal voltage to the local patch of the cell in the patch-clamp mode and detect the corresponding membrane current; A phase demodulation unit is configured to perform phase demodulation of the corresponding membrane current signal based on the phase of the input sinusoidal voltage, obtain the imaginary part signal and the real part signal of the membrane current, and perform real-time calibration and scaling; A membrane capacitance calculation unit is configured to detect and calculate the membrane capacitance change caused by the virus or pseudo-virus infecting the cells based on the current real part signal and the current imaginary part signal based on the calibration and scaling. The event recognition unit is used for recognizing the virus or pseudo-virus infection cell event and its mode according to the dynamic characteristics and polarity of the membrane patch capacitance change, and estimating the size of the virus or pseudo-virus according to the size of the membrane patch capacitance change.

7. The system for detecting viral or pseudo viral infection of cells according to claim 5, wherein, The membrane current detection unit is configured with a phase demodulation module and uses the signal generation function of the phase demodulation module to apply a high-frequency sinusoidal voltage to the attached membrane patch and detect the corresponding membrane current on the basis of realizing the cell-attached membrane patch clamp recording with the giant resistance electrical isolation inside and outside the electrode.

8. The system for detecting viral or pseudo viral infection of cells according to claim 5, wherein, The phase demodulation unit detects the corresponding membrane current after the voltage is applied to the cell membrane patch in the attached membrane patch clamp mode, and performs phase demodulation on the membrane current to obtain the current real part signal and the current imaginary part signal.

9. The system for detecting viral or pseudo viral infection of cells according to claim 5, wherein, The membrane capacitance calculation unit detects and calculates the membrane capacitance change caused by the virus or pseudo-virus infection cell according to the current real part signal and the current imaginary part signal on the basis of calibration and scaling, and calculates the membrane capacitance change according to the current real part and imaginary part signals by calibration to make the changes of the imaginary part signal and the real part signal directly related to the capacitance change, and by scaling to determine the scale coefficient K according to the following calculation formula The membrane capacitance change is calculated as follows: ； Wherein, Cv is the cell membrane capacitance change of the detection area, Re is the change of the current real part signal, and Im is the change of the current imaginary part signal.

10. The system for detecting viral or pseudo viral infection of a cell according to claim 5, wherein: The event recognition unit recognizes the virus or pseudo-virus infection cell event and its mode according to the dynamic characteristics and polarity of the membrane patch capacitance change, and estimates the size of the virus or pseudo-virus according to the size of the membrane patch capacitance change, and the virus or pseudo-virus size is estimated according to the ratio of the size of the membrane patch capacitance change to the unit membrane area capacitance.

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