CN115856271A - Method and system for detecting virus or pseudovirus-infected cells - Google Patents

Abstract

The invention discloses a method for detecting virus or pseudovirus infected cells, which comprises the steps of applying sinusoidal voltage to an attached membrane in an attached membrane clamp mode and detecting corresponding membrane current; inducing virus or pseudovirus to infect cells by combining the virus or pseudovirus in the electrode internal liquid with a receptor on the attached membrane; phase demodulation is carried out on corresponding membrane current signals by taking the phase of input sinusoidal voltage as a standard, so as to obtain the imaginary part and the real part of the membrane current, and real-time calibration and calibration are carried out; calculating the film capacitance of the attached film on the basis of calibration and calibration according to the current real part signal and the current imaginary part signal; and identifying virus or pseudovirus infected cell events and modes thereof according to the dynamic characteristics and polarity of the capacitance change of the diaphragm, and estimating the scale of the virus or pseudovirus according to the size of the capacitance change of the diaphragm. The invention can realize real-time high-precision detection on various viruses or pseudoviruses infecting host cells in a capsule membrane and cell membrane fusion mode and a virus or pseudovirus particle endocytosis mode on acute separated cells, cultured cells and separated tissue living cells.

Description

Method and system for detecting virus or pseudovirus-infected cells

technical field

The invention relates to the technical field of biological and medical detection, in particular to a method and system for detecting cells infected by viruses or pseudoviruses.

Background technique

The research on viruses and the development of therapeutic drugs all start from various links such as virus infection, reverse transcription, replication and self-packaging. Taking the new coronavirus as an example, the first line of defense—the detection and research of cells infected by the new coronavirus is crucial to the prevention and control of the new coronavirus. The diameter of the virus is about tens to hundreds of nanometers, and the process of infecting cells is completed in milliseconds. At present, the observation techniques used for virus detection and infected cells in the field include: electron microscope observation, high-resolution fluorescence microscopy imaging based on protein expression, western blot and nucleic acid detection and other methods.

Among them, the electron microscope observation method has high spatial precision and intuitiveness, and can observe a single virus infection event at a low probability, but this method cannot realize the observation of living cells, and can only complete the observation of cells at a certain time point. or static detection of subcellular morphology. There are limitations such as lengthy and cumbersome processing and easy contamination and damage of samples. High-resolution fluorescence microscopy imaging based on protein expression can detect the movement trajectory of individual virus particles in real time at a lower time resolution (sub-second level), but the observation samples are limited to cultured cells, and the virus must be genetically or proteinically analyzed. remodel. Methods such as immunoblotting and nucleic acid detection have no spatiotemporal information and limited signal accuracy, making it difficult to accurately and real-time monitor the dynamics of virus infection in living cells.

In short, these existing methods are either difficult to accurately and real-time monitor the kinetic process of virus infecting living cells due to technical reasons, or may change the virus traits due to the necessity of gene or protein modification of the virus. In addition, none of the above methods can directly observe virus infection on human living tissues, so that the corresponding observation is still far from the real human pathological phenomenon, which limits the diagnosis and treatment of virus-related diseases and the process of drug development. The above technical problems urgently need to be solved from a new perspective and innovative means to develop a method that can accurately and real-time detect virus infecting living cells.

Contents of the invention

Aiming at the problems existing in the existing virus-infected cell detection technology, the present invention realizes the ability to detect various viruses or pseudoviruses that replace RNA or DNA on living cells in acutely isolated cells, cultured cells, and isolated tissues of animals and humans. Real-time, high-precision detection of infestation.

Technical scheme of the present invention is realized like this:

According to one aspect of the present invention, a method for detecting virus or pseudovirus-infected cells is provided.

The detection method of this virus or pseudovirus infection cell comprises:

Establish an attached patch clamp mode so that the virus or pseudovirus in the electrode fluid can bind to the receptor on the membrane contained in the electrode to induce the virus or pseudovirus to infect cells;

In the attached patch clamp mode, a sinusoidal voltage is applied to the local attached patch of the cell and the corresponding membrane current is detected;

Use the phase of the input sinusoidal voltage as the standard to perform phase detection and demodulation on the corresponding membrane current signal, obtain the imaginary part signal and real part signal of the membrane current, and calibrate and scale in real time;

According to the current real part signal and the current imaginary part signal, on the basis of calibration and calibration, detect and calculate the change of membrane capacitance caused by virus or pseudovirus infecting cells;

According to the dynamic characteristics and polarity of the membrane capacitance change, the virus or pseudovirus infection cell event and its mode are identified, and the scale of the virus or pseudovirus is estimated according to the size of the membrane capacitance change.

In addition, the detection method of the virus or pseudovirus infecting cells also includes: on the basis of realizing the cell-attached patch clamp recording with giant resistance electrical isolation inside and outside the electrode, configuring a phase detection and demodulation module and using its signal generation function Apply a high-frequency sine wave voltage to the attached membrane and detect the corresponding membrane current.

Among them, the phase detection and demodulation of the corresponding membrane current signal is carried out based on the phase of the input sinusoidal voltage, and the imaginary part and the real part of the membrane current are obtained. current, and perform phase detection and demodulation on the membrane current to obtain a current real part signal and a current imaginary part signal of the membrane current.

Among them, according to the current real part signal and the current imaginary part signal, on the basis of calibration and calibration, the detection and calculation of the membrane capacitance change caused by the virus or pseudovirus infection of the cell includes:

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 scaling coefficient K is determined through calibration, and the film capacitance change is calculated by the following calculation formula:

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

Among them, according to the dynamic characteristics and polarity of the membrane capacitance change, the virus or pseudovirus infection cell event and its method include: if the membrane capacitance rises step by step, it is recognized as the invasion of the virus or pseudovirus envelope and the cell membrane fusion; Cell-infecting events; if the membrane capacitance drops step by step, it is recognized as an infecting cell event in the endocytosis of viral particles or pseudoviral particles. Estimate the scale of virus or pseudovirus according to the size of membrane capacitance change.

According to another aspect of the present invention, a detection system for virus or pseudovirus-infected cells is provided.

The detection system of the virus or pseudovirus infecting cells, including:

The infection inducing unit is used to establish an attached patch clamp mode so that the virus or pseudovirus in the electrode inner solution can bind to the receptor on the membrane contained in the electrode to induce the virus or pseudovirus to infect cells;

The membrane current detection unit is used to apply a sinusoidal voltage to the local attached membrane of the cell and detect the corresponding membrane current in the attached patch clamp mode;

The phase detection and demodulation unit is used to perform phase detection and demodulation on the corresponding membrane current signal based on 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;

The membrane capacitance calculation unit is used to detect and calculate the change of membrane capacitance caused by virus or pseudovirus infection on the basis of calibration and calibration according to the current real part signal and the current imaginary part signal;

The event identification unit is used to identify the virus or pseudovirus infection cell event and its mode according to the dynamic characteristics and polarity of the diaphragm capacitance change, and to estimate the scale of the virus or pseudovirus according to the magnitude of the diaphragm capacitance change.

Wherein, the membrane current detection unit, on the basis of realizing the cell-attached patch clamp recording in which the inside and outside of the electrode achieve giant resistance electrical isolation, is equipped with a phase detection demodulation module and uses its signal generation function to apply high frequency to the attached patch. Sine wave voltage and detect the corresponding membrane current.

Wherein, when the phase detection and demodulation unit performs phase detection and demodulation on the corresponding membrane current signal based on the phase of the input sinusoidal voltage to obtain the imaginary part and the real part of the membrane current, the detection is performed in the attached patch clamp mode The corresponding membrane current after the voltage is applied to the cell membrane, and the phase detection and demodulation of the membrane current is performed to obtain the current real part signal and the current imaginary part signal of the membrane current.

Wherein, when the membrane capacitance calculation unit detects and calculates the membrane capacitance change caused by virus or pseudovirus infection on the basis of calibration and calibration according to the current real part signal and the current imaginary part signal, according to the current actual part 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, the scaling coefficient K is determined through calibration, and the film capacitance change is calculated by the following calculation formula:

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

Wherein, when the event recognition unit recognizes the virus or pseudovirus infection cell event and its mode according to the dynamic characteristics and polarity of the membrane capacitance change, if the membrane capacitance rises step by step, it is recognized as a virus or pseudovirus envelope Cell-infecting event in the way of fusion with the cell membrane; if the membrane capacitance drops step by step, it is recognized as a cell-infecting event in the way of endocytosis of viral particles or pseudoviral particles. When estimating the scale of viruses or pseudoviruses according to the size of the change in membrane capacitance, the scale of viruses or pseudoviruses (capsule area or endocytic envelope) is estimated based on the ratio of the size of the change in membrane capacitance to the capacitance per unit membrane area.

Beneficial effect:

The present invention uses electrical methods to realize real-time monitoring of the kinetic process of wild and various mutant viruses or pseudoviruses infecting cells under physiological conditions and pharmacological intervention conditions, and its precision reaches the observation of single virus or pseudoviruses infecting cells; The detection technology can also be used in a high-throughput large-scale rapid drug screening system by expanding the measurement channel and capacity. The detection technology of the present invention can also be used to infect artificial membrane systems, cultured cell systems and human tissue living bodies with viruses or pseudoviruses. The direct and high-precision observation of cells has great potential in virus prevention and treatment.

Compared with the electron microscope imaging method, which cannot realize the observation of living cells, it can only complete the static detection of cells or subcellular morphology at a certain time point, and the processing process is tedious and cumbersome, and the samples are easily contaminated and damaged. Compared with protein expression-based The high-resolution fluorescence microscopy imaging observation samples are limited to cultured cells, and viruses must be genetically or protein modified. Compared with methods such as immunoblotting and nucleic acid detection, there is no temporal and spatial information and the signal accuracy is limited, so it is difficult to accurately and real-time To monitor the limitations of the kinetic process of virus infecting living cells, the present invention can be used in acutely isolated cells, cultured cells, and isolated tissue living cells, especially on isolated cells without sample contamination and damage and without gene or protein modification of the virus. Realize the dynamic, direct and high-precision real-time detection of the dynamic process of virus infecting living cells on living cells of human tissues. It is the observation method of virus infecting living cells closest to the real human pathological conditions so far.

Description of drawings

In order to more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the following will briefly introduce the accompanying drawings required in the embodiments. Obviously, the accompanying drawings in the following description are only some of the present invention. Embodiments, for those of ordinary skill in the art, other drawings can also be obtained based on these drawings without any creative effort.

Fig. 1 is the schematic flow chart of the detection method of the false new coronavirus infection cell of the embodiment of the present invention;

Fig. 2 is the detection system structural diagram of the false new coronavirus infection cell according to the embodiment of the present invention;

Fig. 3 is a flow chart of detection of a single pseudo-new coronavirus infection event in a cell line expressing ACE2/TMPRSS2 according to an embodiment of the present invention;

Fig. 4 is a flow chart of detection of a single pseudo-new coronavirus infection event in HEK293T, Coca-2 and other cell lines transfected with ACE2 and TMPRSS2 according to an embodiment of the present invention;

Fig. 5 is a flow chart of detecting a single pseudo-new coronavirus infection event in alveolar cells in human lung tissue according to an embodiment of the present invention.

Detailed ways

The technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. Apparently, the described embodiments are only some of the embodiments of the present invention, but not all of them. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention belong to the protection scope of the present invention.

According to an embodiment of the present invention, a method and system for detecting pseudo-new coronavirus infecting HEK293T cells expressing ACE2 and TMPRSS2 proteins are provided.

As shown in Figure 1, a method for detecting cells infected with a pseudo-new coronavirus according to an embodiment of the present invention includes:

Step S101, establishing an attached patch clamp mode so that the pseudo-new coronavirus in the electrode inner liquid can bind to the receptor on the membrane contained in the electrode to induce the pseudo-new coronavirus to infect cells;

Step S103, applying an input sinusoidal voltage to the local attached patch of the cell in the attached patch clamp mode and detecting the corresponding membrane current;

Step S105, using the phase of the input sinusoidal voltage as a standard to perform phase detection and demodulation on the corresponding membrane current signal, obtain the imaginary part and real part of the membrane current, and calibrate and scale in real time;

Step S107, according to the current real part signal and the current imaginary part signal, on the basis of calibration and calibration, detect and calculate the membrane capacitance change caused by the fake new coronavirus infected cells;

Step S109, identifying the pseudo-new coronavirus infection cell event and its mode according to the dynamic characteristics and polarity of the diaphragm capacitance change, and estimating the scale of the pseudo-new coronavirus according to the magnitude of the diaphragm capacitance change.

In one embodiment, the method for detecting cells infected by the pseudo-new coronavirus further includes: by configuring the phase detection and demodulation module, using the signal generation function of the lock-in amplifier to apply a sinusoidal voltage to the diaphragm contained in the electrode; In the attached patch clamp mode, the membrane current induced by applying voltage to the diaphragm is used. The phase detection function of the lock-in amplifier is used to demodulate the corresponding membrane current signal based on the phase of the input sinusoidal voltage, and the virtual membrane current is obtained. department and real department;

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

Among them, Cv is the change of the cell membrane capacitance in the detection area, Re is the change of the real part signal of the current, and Im is the change of the imaginary part signal of the current.

In one embodiment, the virus or pseudovirus infection event identification and analysis module identifies the pseudo-new coronavirus infection cell event and its mode according to the dynamic characteristics and polarity of the diaphragm capacitance change, and according to the magnitude of the diaphragm capacitance change Estimate the scale of the pseudo-new coronavirus, including: if the membrane capacitance rises step by step, it will be recognized as the infecting cell event of the fusion of the pseudo-new coronavirus capsule membrane and the cell membrane; Infected cells by endocytosis. The scale of the pseudo-new coronavirus (capsule area or endocytic envelope) is estimated based on the ratio of the change in membrane capacitance to the capacitance per unit membrane area.

In order to facilitate the understanding of the above-mentioned technical solution of the present invention, the above-mentioned technical solution of the present invention will be described in detail below through specific application descriptions and experiments.

In an example of detection of pseudo-new coronavirus infecting cultured cells expressing the new coronavirus receptor protein ACE2 and auxiliary protein TMPRSS2, the detection experimental process of pseudo-new coronavirus infecting the cultured cells is shown in Figure 3: In HEK293T cells Express the new coronavirus receptor protein ACE2 and the auxiliary protein TMPRSS2 as candidate host cells. During the experiment, the candidate host cells (groups) were moved into the recording tank containing extracellular fluid in the patch clamp electrophysiological platform.

Wherein the formula of extracellular fluid is as follows:

Element g/L NaCl 9 KCl 0.4 MgCl2 0.5 CaCl2 0.3 HEPES 2.5 D-Glucose 2 distilled water Add to 1000ml, adjust pH to 7.4

The detection activity is carried out in the cell-attached patch clamp mode: the glass microelectrode contains the test solution of pseudo-new coronavirus, the tip diameter is about 1 μm, and it is clamped on the conventional electrophysiological amplifier probe, and the negative pressure airflow is applied to make the tip contact with the cell membrane The surface is tightly sealed to realize the electrical isolation of giant resistance inside and outside the electrode to greatly reduce background noise and achieve high signal-to-noise ratio measurement. The detection system for cells infected by the pseudo-new coronavirus is shown in Figure 2. 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 module. Cell event recognition and analysis module composition. Specifically include:

Electrophysiological signal acquisition module: with the patch clamp amplifier as the main body, it realizes the cell-attached patch clamp recording with giant resistance electrical isolation inside and outside the electrode, and collects the attached patch in the high-frequency sine wave voltage clamp mode modulated by DC level The current is amplified with high gain and then output to the phase detection demodulation module.

Phase detection and demodulation module: output the high-frequency sine wave (non-RF sine wave above 5kHz) voltage as a modulation signal to the electrophysiological signal acquisition module, and use this as a phase-locking standard to perform a phase-locking standard on the current signal input from the electrophysiological signal acquisition module Phase detection demodulation to obtain the imaginary part and real part of the signal;

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

Membrane capacitance signal calibration module: used for calibration of the detected membrane capacitance signal. The scaling factor K is determined by scaling.

Membrane capacitance detection module: use a high-speed analog/digital converter to convert the imaginary part and real part signals obtained by phase detection and demodulation of the attached diaphragm current signal output by the phase detection demodulation module to realize analog-to-digital conversion, which is realized by combining software and hardware Capacitance and conductance signal detection of the attached diaphragm based on real-time phase calibration and signal scaling, and the corresponding information is stored in the computer.

Recognition and analysis module of virus or pseudovirus infecting cell events: use software to post-process the attached membrane capacitance and conductance signals stored in the computer. According to the dynamic characteristics and polarity of the membrane capacitance change, when identifying the pseudo-new coronavirus infection cell event and its mode, if the membrane capacitance rises step by step, it is recognized as the infection cell event of the fusion method of the pseudo-new coronavirus capsule membrane and the cell membrane; If the membrane capacitance drops step by step, it is recognized as an infecting cell event of the pseudo-new coronavirus particle endocytosis. When estimating the scale of the pseudo-new coronavirus based on the change in membrane capacitance, the scale of the pseudo-new coronavirus (capsule area or endocytic envelope) is estimated based on the ratio of the change in membrane capacitance to the capacitance per unit membrane area. In this way, the size, infection mode, dynamic process, and timing characteristics of the pseudo-new coronavirus were analyzed.

When it is specifically applied to different scenarios, taking fake new coronavirus as an example, the experimental process of using the detection system of the present invention to complete the detection of fake new coronavirus infected cells is as follows:

1) Artificial cell membrane

As shown in Figure 3, for liposomes embedded in the ACE2/TMPRSS2 membrane protein, cell-attached lower membrane capacitance measurement technology was used to measure and detect pseudo-new coronavirus infection cell events.

2) Cultured cell lines

As shown in Figure 4, HEK293T, Coca-2 and other cell lines were transfected with ACE2 and TMPRSS2, and cell-attached lower membrane capacitance measurement technology was used to detect pseudo-new coronavirus-infected cells.

Preparation of human lung tissue biopsy and detection of pseudo-new coronavirus infection.

As shown in Figure 5, the living pathological tissue of the patient's lung surgically removed was used as a sample, and the sample was prepared into a living lung tissue slice with a thickness of 150-400 microns using a vibrating microtome, and then the cell-attached membrane capacitance was made on the alveolar epithelial cells Detection, using membrane capacitance measurement technology to detect a single pseudo-new coronavirus-infected cell event.

It can be seen that, with the help of the above technical scheme, the present invention uses electrical methods to treat pseudo-new coronaviruses in the form of capsule and cell membrane fusion and pseudo-new coronavirus particles at the level of artificial cell membranes, animal and human isolated cells, cultured cells, and living tissues. Real-time detection of host cells infected by endocytosis, the accuracy of which can reach the observation of a single pseudo-new coronavirus infection cell event.

The detection technology of the present invention can also be used for direct and high-precision observation of other viruses or pseudoviruses infecting artificial membrane systems, cultured cell systems, and living cells of human tissues. The detection technology can also be used for high Throughput large-scale rapid drug screening has great potential in virus prevention and treatment.

The above descriptions are only preferred embodiments of the present invention, and are not intended to limit the present invention. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principles of the present invention shall be included in the scope of the present invention. within the scope of protection.

Claims (10)

1. A detection method for virus or pseudovirus-infected cells, characterized in that, comprising: Establish an attached patch clamp mode so that the virus or pseudovirus in the electrode fluid can bind to the receptor on the membrane contained in the electrode to induce the virus or pseudovirus to infect cells; In the attached patch clamp mode, a sinusoidal voltage is applied to the local attached patch of the cell and the corresponding membrane current is detected; Use the phase of the input sinusoidal voltage as the standard to perform phase detection and demodulation on the corresponding membrane current signal, obtain the imaginary part signal and real part signal of the membrane current, and calibrate and scale in real time; According to the current real part signal and the current imaginary part signal, on the basis of calibration and calibration, detect and calculate the change of membrane capacitance caused by virus or pseudovirus infecting cells; According to the dynamic characteristics and polarity of the membrane capacitance change, the virus or pseudovirus infection cell event and its mode are identified, and the scale of the virus or pseudovirus is estimated according to the size of the membrane capacitance change. 2. the detection method of virus or pseudovirus infecting cells according to claim 1, is characterized in that, also comprises: on the basis of realizing the cell-attached patch-clamp recording that reaches giant resistance electrical isolation inside and outside the electrode, configure The phase detection and demodulation module uses its signal generation function to apply high-frequency sine wave voltage to the attached diaphragm and detect the corresponding membrane current. 3. the detection method of virus or pseudovirus infection cell according to claim 1, it is characterized in that, take the phase of input sinusoidal voltage as a standard to carry out phase detection demodulation to corresponding membrane current signal, obtain the imaginary part of membrane current and The real part includes: detecting the corresponding membrane current after the voltage is applied to the cell membrane in the attached patch clamp mode, and performing phase detection and demodulation on the membrane current to obtain the current real part signal and the current imaginary part of the membrane current Signal. 4. the detection method of virus according to claim 1 or pseudovirus infecting cell is characterized in that, detects and calculates virus or pseudovirus invasion on the basis of calibration and calibration according to current real part signal and current imaginary part signal Changes in membrane capacitance caused by stained cells include: 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 scaling coefficient K is determined through calibration, and the film capacitance change is calculated by the following calculation formula:

Among them, Cv is the change of the cell membrane capacitance in the detection area, Re is the change of the real part signal of the current, and Im is the change of the imaginary part signal of the current. 5. the detection method of virus or pseudovirus infecting cells according to claim 1, is characterized in that, according to the kinetic characteristic and the polarity identification virus of diaphragm capacitance change or pseudovirus infecting cell event and its mode, according to The size of the diaphragm capacitance change estimates the scale of the virus or pseudovirus, including: If the membrane capacitance rises step by step, it is recognized as an infecting cell event in which the virus or pseudovirus envelope fuses with the cell membrane; If the membrane capacitance drops step by step, it is recognized as an infecting cell event in the endocytosis of virus particles or pseudovirus particles; Virus or pseudovirus (envelope area or endocytic envelope) scale is estimated from the ratio of the change in membrane capacitance to the capacitance per unit membrane area. 6. A detection system for virus or pseudovirus-infected cells, characterized in that, comprising: The infection inducing unit is used to establish an attached patch clamp mode so that the virus or pseudovirus in the electrode inner solution can bind to the receptor on the membrane contained in the electrode to induce the virus or pseudovirus to infect cells; The membrane current detection unit is used to apply a sinusoidal voltage to the local attached membrane of the cell and detect the corresponding membrane current in the attached patch clamp mode; The phase detection and demodulation unit is used to perform phase detection and demodulation on the corresponding membrane current signal based on 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; The membrane capacitance calculation unit is used to detect and calculate the change of membrane capacitance caused by virus or pseudovirus infection on the basis of calibration and calibration according to the current real part signal and the current imaginary part signal; The event recognition unit is used to identify the virus or pseudovirus infection cell event and its mode according to the dynamic characteristics and polarity of the diaphragm capacitance change, and to estimate the scale of the virus or pseudovirus according to the magnitude of the diaphragm capacitance change. 7. the detection system of virus or pseudovirus infecting cells according to claim 6, is characterized in that, described membrane electric current detection unit reaches the basis of the cell-attached patch clamp recording of giant resistance electrical isolation inside and outside the realization electrode Above, configure the phase detection and demodulation module and use its signal generation function to apply high-frequency sine wave voltage to the attached diaphragm and detect the corresponding membrane current. 8. The detection system for virus or pseudovirus-infected cells according to claim 6, characterized in that, the phase detection and demodulation unit carries out phase detection and demodulation to the corresponding membrane current signal based on the phase of the input sinusoidal voltage , to obtain the imaginary part and real part of the membrane current, detect the corresponding membrane current after the voltage is applied to the cell membrane in the attached patch clamp mode, and perform phase detection and demodulation on the membrane current to obtain the membrane current Current real part signal and current imaginary part signal. 9. The detection system of virus or pseudovirus infecting cells according to claim 6, characterized in that, said membrane capacitance calculation unit detects on the basis of calibration and scaling according to current real part signal and current imaginary part signal And when calculating the change of membrane capacitance caused by virus or pseudovirus infecting cells, according to the real part and imaginary part signals of the current, the changes of the imaginary part signal and the real part signal are directly related to the capacitance change through calibration, and determined by calibration Scale factor K, the film capacitance change is calculated by the following formula:

Among them, Cv is the change of the cell membrane capacitance in the detection area, Re is the change of the real part signal of the current, and Im is the change of the imaginary part signal of the current. 10. The detection system of virus or pseudovirus-infected cells according to claim 6, characterized in that: said event recognition unit recognizes virus or pseudovirus-infected cells according to the kinetic characteristics and polarity of the diaphragm capacitance change In the case of events and their methods, if the membrane capacitance rises step by step, it is recognized as an infecting cell event in the form of fusion between the virus or pseudovirus envelope and the cell membrane; if the membrane capacitance decreases step by step, it is recognized as endocytosis of virus particles or pseudovirus particles In the event of infecting cells of the mode, when estimating the scale of the virus or pseudovirus according to the size of the membrane capacitance change, the scale of the virus or pseudovirus (capsule area or endocytic envelope) is based on the size of the membrane capacitance change and the unit membrane area capacitance ratio to estimate.

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