WO2018170875A1 - Pet disease prevention and diagnosis method based on nanometer-scale gene anchoring and microfluidic immunological agglutination technology - Google Patents

Abstract

Provided is a pet disease prevention and diagnosis method based on a nanometer-scale gene anchoring and microfluidic immunological agglutination technology, comprising: a microfluidic control chip, a photoelectric detector, a wireless transmission module, an interface device card inserted into a microcomputer PCI slot, and a software measurement and control module inside the microcomputer; and further comprising: a DNA sample preparation module, magnetic beads for surface modification ligands, a DNA library chip, a sequence signal collection and analyzing module, and a data display module. Compared with the separate use of PCT and CRP, a combined application thereof provides a wider range of use, greater convenience, and more accurate diagnosis results. In addition, comprehensive diagnosis is performed on bacterial infections, and prevention and diagnosis are performed on hereditary diseases and infectious diseases of pets by using a gene sequencing technology, such that pets can be treated in advance of symptoms, treatment timing is controlled, and pet health is ensured.

Description

Pet anti-diagnosis method for gene nano anchoring and microfluidic immunoagglutination technology Technical field

The invention relates to the technical field of pet prevention and diagnosis, in particular to a pet anti-diagnosis method of gene nano anchoring and microfluidic immunoagglutination technology.

Background technique

With the development of social economy and the acceleration of urbanization, people's spiritual and material life have been continuously improved, the pace of social aging has accelerated, and the objective factors such as the popularity of only-child families and Dink families have emerged. People's leisure, consumption and Emotional support methods are also diversified. Pets have entered the homes of ordinary people, and keeping pets is an indispensable pastime for families.

Pet breeding is not only related to the development of human society, but also related to economic development. Over the past 30 years of reform and opening up, China's GDP has developed at a rate of nearly 10% per year. The progress of urbanization has reached more than 50%. With the result, the size of the family has shrunk, the number of families has increased, and the family structure has taken place. Change; the problem of population aging has become increasingly prominent, and the situation of empty nesters and young people has made the urban residents' emotional support, leisure and entertainment, and consumer spending also diversified. Because "to some extent, animals are our tools to carry ideas," keeping pets has gradually become one of the ways in which young people show their individuality and the feelings of the elderly, and pets have become important members of the family.

With the development of the pet industry, the concept of pet breeding has gradually changed. Pets have become an indispensable part of the family, and the related pet prevention and diagnosis industry has undergone major changes, and the disease has become more complicated, preventive and The diversified diagnostic tools and more humane services have brought unprecedented opportunities and challenges to the entire pet prevention and diagnostic industry.

Inherited errors of disease (IMD), also known as inborn errors of metabolism (IEM), are codes of certain enzymes, transporters, membranes, or receptors necessary to maintain normal metabolism in the body. A mutation in a gene that causes a change in the function of the encoded product, and a corresponding type of disease in which laboratory abnormalities and clinical symptoms occur. It involves the abnormal metabolism of various substances such as amino acids, organic acids, fatty acids, urea cycle, carbohydrates, lysosomes, and steroids. Most of these diseases are monogenic genetic diseases. Although the single disease has a low prevalence rate, it has a wide variety, so the overall incidence rate is high, and the overall incidence rate is about 1/800.

In recent years, viral infectious diseases have become more and more serious, and the infection route is complicated, and the pathogen variability is large, which undoubtedly brings great challenges to clinical testing. Traditional pathogen detection methods such as clinical symptoms, pathological changes, virus isolation, artificial pathogenic tests, etc. to diagnose pathogens, serum separation, electron microscopy, etc., large workload, complex technology, long time, is not suitable for effective monitoring of infectious diseases And prevention and control. This requires a fast and flexible method to detect pathogenic nucleic acids to tightly control the occurrence of infectious diseases.

C reactive protein (CRP) is synthesized by the liver and is a typical acute phase protein. Its content in normal serum is extremely small. It is in the plasma of patients with acute inflammation, malignant tumor, ischemia, tissue damage and other diseases. Among them, the CRP content can be increased by a thousand times. It increased in the early 4h-6h of inflammation and reached a peak in 36h-50h. Once the infection is controlled, its content will quickly return to normal levels. Studies have found that bacterial infections can cause elevated levels of CRP, and when more than 140 mg/L, the presence of bacterial infections can be determined. And its level is positively correlated with the degree of infection; when the virus is infected, the CRP level is generally not elevated. For the diagnosis of acute infectious diseases, the detection of serum CRP is more sensitive, more accurate and more reliable than the total number of white blood cells.

Procalcitonin (PCT) is a precursor of calcitonin, which is a glycoprotein with a half-life of 25h to 30h, and has good stability in vitro and in vivo. It is produced by C cells of the thyroid gland under physiological conditions, but its content is extremely low (less than 0.1 μg/L). It is considered abnormal when its content is more than 0.5 μg/L. The study found that PCT is a diagnostic indicator for early bacterial or fungal infections, which is elevated at the time of infection. The level of non-infection is generally not elevated, and the magnitude of the increase is closely related to the severity of the infection. A comparative analysis of bacterial infections and viral infections revealed that serum PCT levels were significantly elevated in bacterial infections, whereas serum PCT levels were not significantly elevated in viral infections. A comparative analysis of CRP and PCT found that the increase in PCT was earlier than the increase in CRP. In local infections, PCT generally does not increase, but CRP increases. These studies indicate that CRP is an important predictor of non-systemic infection; PCT is a better indicator of systemic infection.

Summary of the invention

The object of the present invention is to provide a pet anti-diagnosis method for gene nano-anchoring and microfluidic immunoagglutination technology, which utilizes gene nano-anchoring technology to prevent and diagnose pet genetic diseases and viral infections, and can be more timely and effective for pets. Treatment will improve the cure rate of pets to a greater extent and ensure the health of pets. At the same time, through the combined application of PCT and CRP, it will be more widely used and more convenient when used alone, and the diagnosis results will be more accurate.

The invention provides a pet anti-diagnosis method for gene nano anchoring and microfluidic immunoagglutination technology, comprising: a microfluidic chip, a photodetector, a wireless transmission module, an interface device card inserted into a PCI PCI slot, located inside the microcomputer Software measurement and control module; further comprises a DNA sample preparation module, a surface modified ligand magnetic bead, a DNA library chip, a sequence signal acquisition and analysis module, a data display module; wherein the microfluidic chip comprises a sample injection zone, a dyeing zone, a reaction zone, CRP and PCT antibodies are respectively immobilized in the reaction zone, wherein the photodetector transmits a signal generated by the antigen-antibody agglutination reaction on the microfluidic chip to the wireless transmission module, and the wireless transmission module transmits the signal to the interface device through wireless transmission. The card, the interface device card is connected to the communication between the computer and the detector, and the analog signal generated by the photodetector can be collected and transmitted to the software measurement and control module by inserting in the PCI card slot, and the software measurement and control module includes A/D conversion, Digital signal analysis processing, D/A conversion; said A/D conversion: converting two analog signals into digital signals The digital signal analysis process: analyzing the digital signal, treating the signal and its noise as a random signal, and using the statistical features, reducing the noise to obtain the signal itself and integrating the signal to obtain the best digital signal, the D /A conversion: converts the best digital signal analyzed and converted into the corresponding analog signal; wherein the magnetic beads of the surface modification ligand have the same polarity, and the magnetic beads mutually repel each other, and the DNA library chip has a surface modification ligand The magnetic beads match the active sites, the active sites have opposite polarities to the magnetic beads, and are adsorbed by the magnetic beads. The sequence signal acquisition and analysis module collects and analyzes the optical signals, and the optical signals are digitally processed and then displayed through the data. The module obtains the sequence of the gene to be tested.

Further, the microfluidic chip is made of glass, and the microchannel is formed by soft etching, and the CRP antibody and the PCT antibody are preliminarily fixed on the aldehyde-treated glass slide at a predetermined position and then at 37. Incubate for 1 hour at °C.

Further, the digital signal analysis processing uses digital filtering, FFT analysis, single product extraction, single frequency detection, wavelet denoising, autocorrelation analysis, weighted average filtering, anti-pulse interference average filtering, and program judgment filtering. , adaptive filtering, linear neural network denoising, curve fitting methods to achieve related functions.

Further, the software measurement and control module performs A/D conversion on the input analog signal by starting digital signal analysis processing, and each When a data is collected, it is sent to the digital signal analysis processing for filtering operation, and the operation result is converted into analog quantity by D/A conversion. The above process is continuously repeated, and the filtered analog signal is obtained at the output of the D/A conversion. In the operation process, not only the signal is denoised, but also the two analog signals outputted by the series module of the conductivity cell are converted into two digital signals by A/D conversion, and further analyzed and sent to D/A. The conversion is converted to an analog quantity, which in turn further forms the data.

Further, the DNA library chip comprises a pet genetic disease gene chip and a pet susceptible virus gene chip.

Further, the pet susceptible virus microarray has a DNA probe with different viruses at each active site, and the pet is infected or infected with a virus, and then further determined by gene sequencing. Determine if you are infected with a virus.

The beneficial effects of the invention are that, through the combined application of PCT and CRP, the application range is wider and more convenient than that of the two, and the diagnosis result is more accurate, and the bacterial infection is comprehensively diagnosed, and the gene sequencing is utilized. The technology prevents and diagnoses pet's hereditary diseases and infectious diseases. It can treat pets in advance, grasp the timing of treatment, and better ensure the health of pets.

DRAWINGS

Figure 1 is a schematic view of the structure of the present invention

detailed description

The technical solutions in the embodiments of the present invention are clearly and completely described below. It is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without creative efforts are within the scope of the present invention.

The invention provides a pet anti-diagnosis method for gene nano anchoring and microfluidic immunoagglutination technology, comprising: a microfluidic chip, a photodetector, a wireless transmission module, an interface device card inserted into a PCI PCI slot, located inside the microcomputer Software measurement and control module; further comprises a DNA sample preparation module, a surface modified ligand magnetic bead, a DNA library chip, a sequence signal acquisition and analysis module, a data display module; wherein the microfluidic chip comprises a sample injection zone, a dyeing zone, a reaction zone, CRP and PCT antibodies are respectively immobilized in the reaction zone, wherein the photodetector transmits a signal generated by the antigen-antibody agglutination reaction on the microfluidic chip to the wireless transmission module, and the wireless transmission module transmits the signal to the interface device through wireless transmission. The card, the interface device card is connected to the communication between the computer and the detector, and the analog signal generated by the photodetector can be collected and transmitted to the software measurement and control module by inserting in the PCI card slot, and the software measurement and control module includes A/D conversion, Digital signal analysis processing, D/A conversion; said A/D conversion: converting two analog signals into digital signals The digital signal analysis process: analyzing the digital signal, treating the signal and its noise as a random signal, and using the statistical features, reducing the noise to obtain the signal itself and integrating the signal to obtain the best digital signal, the D /A conversion: converts the best digital signal analyzed and converted into the corresponding analog signal; wherein the magnetic beads of the surface modification ligand have the same polarity, and the magnetic beads mutually repel each other, and the DNA library chip has a surface modification ligand The magnetic beads match the active sites, the active sites have opposite polarities to the magnetic beads, and are adsorbed by the magnetic beads. The sequence signal acquisition and analysis module collects and analyzes the optical signals, and the optical signals are digitally processed and then displayed through the data. The module obtains the sequence of the gene to be tested.

Further, the microfluidic chip is made of glass, and the microchannel is formed by soft etching, and the CRP antibody and the PCT antibody are preliminarily fixed on the aldehyde-treated glass slide at a predetermined position and then at 37. Incubate for 1 hour at °C.

Further, the digital signal analysis processing uses digital filtering, FFT analysis, single product extraction, single frequency detection, wavelet denoising, autocorrelation analysis, weighted average filtering, anti-pulse interference average filtering, and program judgment filtering. , adaptive filtering, linear neural network denoising, curve fitting methods to achieve related functions.

Further, the software measurement and control module performs A/D conversion on the input analog signal by starting digital signal analysis processing, and each time a data is collected, it is sent to the digital signal analysis processing for filtering operation, and the operation result is sent to D/A conversion. Convert to analog. The above process is continuously repeated, and the filtered analog signal is obtained at the output of the D/A conversion. In the operation process, not only the signal is denoised, but also the two analog signals outputted by the series module of the conductivity cell are converted into two digital signals by A/D conversion, and further analyzed and sent to D/A. The conversion is converted to an analog quantity, which in turn further forms the data.

Further, the DNA library chip comprises a pet genetic disease gene chip and a pet susceptible virus gene chip.

Further, the pet susceptible virus microarray has a DNA probe with different viruses at each active site, and the pet is infected or infected with a virus, and then further determined by gene sequencing. Determine if you are infected with a virus.

Working process: First, the blood sample is added to the system, a part of the blood enters the microfluidic chip injection area, flows through the microchannel to the dyeing zone, and is mixed and incubated with the fluorescent dyeing solution of the dyeing zone, fully combined, and then passed through the microchannel. Further entering the reaction zone, respectively, agglutination reaction with CRP antibody and PCT antibody, after the reaction is completed, using a photodetector for detection, the detection signal is transmitted to the interface device card located in the microcomputer through the wireless transmission module, and inserted into the PCI card The slot can collect the analog signal generated by the photodetector, transmit it to the software measurement and control module, and obtain the required data through the software measurement and control module; another part of the blood enters the DNA sample preparation module, and after the DNA sample is prepared, the surface modification ligand is prepared. The magnetic beads adsorb and adsorb the magnetic beads of the DNA sample into the DNA library chip, and combine with the active site, and the sequence signal acquisition and analysis module collects and analyzes the optical signal, and the optical signal is digitally processed and then obtained through the data display module. The gene sequence to be tested.

Claims (6)

A pet anti-diagnosis method for gene nano-anchoring and microfluidic immunoagglutination technology, comprising: a microfluidic chip, a photodetector, a wireless transmission module, an interface device card inserted into a PCI PCI slot, and software measurement and control located inside the microcomputer The module further comprises a DNA sample preparation module, a surface modified ligand magnetic bead, a DNA library chip, a sequence signal acquisition and analysis module, and a data display module; wherein the microfluidic chip comprises a sample injection zone, a dyeing zone, a reaction zone, and a reaction zone. CRP and PCT antibodies are respectively immobilized, wherein the photodetector transmits a signal generated by the antigen-antibody agglutination reaction on the microfluidic chip to the wireless transmission module, and the wireless transmission module transmits the signal to the interface device card through wireless transmission, and the interface The device card connects the communication between the computer and the detector, and the analog signal generated by the photodetector can be collected and transmitted to the software measurement and control module by inserting in the PCI card slot, and the software measurement and control module includes A/D conversion and digital signal analysis. Processing, D/A conversion; said A/D conversion: converting two analog signals into digital signals; Word signal analysis processing: analyzing the digital signal, treating the signal and its noise as a random signal, by using its statistical characteristics, reducing the noise to obtain the signal itself and integrating the signal to obtain the best digital signal, the D/A conversion: The best digital signal analyzed and transformed into the corresponding analog signal; wherein the magnetic beads of the surface modification ligand have the same polarity, and the magnetic beads mutually repel each other, and the DNA library chip has the magnetic beads matching the surface modification ligand. The active site has an opposite polarity to the magnetic bead, and is adsorbed with the magnetic beads. The sequence signal acquisition and analysis module collects and analyzes the optical signal, and the optical signal is digitally processed and then obtained through the data display module. gene sequence. The pet anti-diagnosis method according to claim 1, wherein the microfluidic chip is made of glass, and the microchannel is formed by soft etching, and the CRP antibody and the PCT antibody are preliminarily fixed to the aldehyde group. The glass slides were treated at predetermined locations and then incubated for 1 hour at 37 °C. The pet anti-diagnosis method according to claim 1, wherein said digital signal analysis processing uses digital filtering, FFT analysis, single product extraction, single frequency detection, wavelet denoising, autocorrelation analysis, weighted average Filtering, anti-pulse interference average filtering, program judgment filtering, adaptive filtering, linear neural network denoising, curve fitting methods to achieve related functions. The pet anti-diagnosis method according to claim 1, wherein the software measurement and control module activates the digital signal The number analysis processing performs A/D conversion on the input analog signal, and each time a data is acquired, it is sent to the digital signal analysis processing for filtering operation, and the operation result is sent to D/A conversion to analog quantity. The above process is continuously repeated, and the filtered analog signal is obtained at the output of the D/A conversion. In the operation process, not only the signal is denoised, but also the two analog signals outputted by the series module of the conductivity cell are converted into two digital signals by A/D conversion, and further analyzed and sent to D/A. The conversion is converted to an analog quantity, which in turn further forms the data. The pet anti-diagnosis method according to claim 1, wherein the DNA library chip comprises a pet genetic disease gene chip and a pet susceptible virus gene chip. The pet anti-diagnosis method according to claim 5, wherein the pet susceptible virus microarray has DNA probes of different viruses at each active site, and whether the pet is infected or infected The virus is initially determined, and then genetic sequencing is used to further determine whether the virus is infected.

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