A solid-state fermentation sample microbial rapid detection kit and a solid-state fermentation sample microbial rapid detection method

Abstract

The present invention discloses a solid-state fermentation sample microorganism rapid detection kit and a solid-state fermentation sample microorganism rapid detection method, and belongs to the field of solid-state fermentation sample microorganism detection. The solid-state fermentation sample microorganism rapid detection kit includes a separation unit for separating microorganisms from a solid-state fermentation sample, a dilution unit for diluting the separated microorganism suspension with a thickener solution, and a chip for incubating and culturing the diluted microorganism suspension so that the microorganism single cell grows into a cell cluster and then performs identification and statistics. The present invention is applied to the solid-state fermentation sample microorganism detection, and has the characteristics of high detection flux, short detection time, visible detection process, and accurate detection results.

Description

Technical Field

[0001] The invention belongs to the field of solid-state fermentation sample microorganism detection, and in particular relates to a solid-state fermentation sample microorganism rapid detection kit and a solid-state fermentation sample microorganism rapid detection method. Background Art

[0002] In the detection of solid-state fermentation samples, we usually focus on indicators such as core microorganisms, flavor substance content, and physical and chemical properties. For the detection of core microorganisms, most of the current methods use qPCR amplification to count the number of core microorganisms. However, the qPCR amplification method has a long detection cycle, is expensive, and is difficult to analyze microbial activity. The use of traditional dilution plate method or identification culture medium to identify and count microorganisms in solid-state fermentation samples has the disadvantages of long detection cycle, low separation efficiency of core microorganisms and low accuracy of species identification, and wastes manpower and material resources.

[0003] In addition, Raman technology is also a commonly used method for microbial detection, but the current Raman detection method has high requirements for samples, usually requiring the sample particle size to be at the micron level, and there are also restrictions on sample types and buffer types. For complex environmental samples, such as solid-state fermentation samples, pretreatment requirements are strict, and the modeling process is complex, and there are problems with low detection throughput and detection accuracy.

[0004] In summary, existing microbial detection technologies such as qPCR and identification culture medium have problems such as long detection cycle, high price, low separation efficiency and low species identification accuracy when facing solid-state fermentation sample detection. When using Raman technology for microbial detection, there are complex sample pretreatment and high restrictions on sample particle size, microbial species and buffer type. Therefore, a solid-state fermentation sample microbial rapid detection kit and a solid-state fermentation sample microbial rapid detection method are needed. Summary of the invention

[0005] In view of the above-mentioned shortcomings of the existing solid-state fermentation sample detection technology, the technical problem to be solved by the present invention is to overcome the problems of long detection cycle and low detection accuracy in the existing solid-state fermentation sample detection, and to propose a solid-state fermentation sample microbial rapid detection kit and a solid-state fermentation sample microbial rapid detection method that can greatly shorten the detection cycle and realize rapid identification and counting of microorganisms in samples.

[0006] In order to solve the technical problem, the technical solution adopted by the present invention is:

[0007] The present invention provides a solid-state fermentation sample microorganism rapid detection kit, comprising a separation unit for separating microorganisms from the solid-state fermentation sample, a dilution unit for diluting the separated microorganism suspension with a thickener solution, and a chip for incubating and culturing the diluted microorganism suspension so that microorganism single cells grow into cell clusters and then performing identification and statistics; the separation unit comprises: a filter element, which is coated on the outside of the solid-state fermentation sample and has a plurality of filter holes; a separation element, in which a solution is provided in a cavity, and the filter element is immersed in the solution; an oscillating element, which is used to oscillate the separation element so that the microorganisms in the solid-state fermentation sample coated in the filter element flow out of the filter element from the filter holes and enter the separation element; The chip comprises: a channel, in which the diluted microbial suspension is incubated and cultured until the single microbial cells grow into cell clusters, and then identification and statistics are performed, and the width-to-height ratio of the dimensions in the channel is less than 40:1; a liquid storage tank, connected to the channel and used to input the diluted microbial suspension into the channel, and the inner cross-sectional width of the liquid storage tank is greater than the droplet width of the diluted microbial suspension; the amount of the diluted microbial suspension added to the channel is controlled to be 80-120% of the channel volume, and when the amount of the diluted microbial suspension added to the channel is greater than the channel volume, the area of ​​the diluted microbial suspension exceeding the channel on the bottom surface of the liquid storage tank does not exceed 60% of the bottom surface area of ​​the liquid storage tank.

[0008] Preferably, the chip further comprises: an upper layer of glass, a channel structure layer, and a bottom layer of glass arranged in sequence from top to bottom; the upper layer of glass, the hollow portion of the channel structure layer, and the bottom layer of glass form a channel; and the liquid storage tank is connected to at least one end of the channel through a through hole.

[0009] Preferably, the liquid storage tanks are arranged in parallel along the left and right edges of the upper glass, and both ends of the channel are connected to the liquid storage tanks arranged at both ends through through holes.

[0010] Preferably, the chip incubates and cultures the diluted microbial suspension in the channel under a constant humidity environment; or after the diluted microbial suspension is injected into the channel, the liquid storage tank is blocked to isolate the channel from the outside.

[0011] Preferably, a plugging agent, a stopper, or a membrane is used to seal the liquid storage tank to isolate the channel from the outside.

[0012] Preferably, the separated microbial suspension is diluted with a thickener solution to a concentration of 1% to 99% of the original microbial suspension.

[0013] Preferably, the filter element includes but is not limited to a filter bag, a filter membrane or a filter screen; the separation element includes but is not limited to a centrifuge tube, a test tube, a conical flask; the solution includes but is not limited to a culture medium and a buffer solution.

[0014] Preferably, a hollow structure is provided on the channel structure layer between adjacent channels for focusing the microscopic field of view.

[0015] The present invention also provides a method for rapid detection of microorganisms in solid-state fermentation samples, which uses the rapid detection kit for microorganisms in solid-state fermentation samples described in any of the above technical solutions to detect the microorganisms in solid-state fermentation samples.

[0016] Preferably, the solid fermentation sample is separated into microorganisms by a separation unit, and the microbial suspension is diluted with a thickener solution, and the diluted microbial suspension is input into the channel by a liquid storage tank for incubation and culture so that the microbial single cells grow into cell clusters for identification and counting.

[0017] Compared with the prior art, the present invention has the following beneficial effects:

[0018] The present invention provides a solid-state fermentation sample microorganism rapid detection kit, which utilizes a separation unit to realize rapid separation of microorganisms and large-particle impurities, thereby realizing rapid pretreatment of the solid-state fermentation sample, extracting the microorganisms therein, reducing the complexity of the solid-state fermentation sample pretreatment, and shortening the pretreatment time; utilizing a dilution unit to dilute the microorganism suspension, thereby reducing the influence of impurities with smaller particle sizes on the process of identifying cell clusters, and at the same time limiting the structure of the chip, ensuring that there are no bubbles in the microorganism suspension in the channel, ensuring the stability of the microorganism suspension in the channel, realizing static culture of the microorganisms in the chip channel, proliferating cells will stay and attach to the mother cell enrichment, while the cells that have not proliferated still maintain a single cell state, and then can distinguish dead cells from living cells by whether they can generate cell clusters, and through the generated cell clusters, the difference between different types of cells is amplified, and different types of cells are distinguished, thereby realizing rapid identification and counting of microorganisms in the sample, and greatly shortening the detection cycle. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 A schematic diagram of the structure of a chip provided by an embodiment of the present invention;

[0020] Figure 2 The variation of the number of acetic acid bacteria with the fermentation days provided in Example 1 of the present invention;

[0021] Figure 3 The activity of yeast A provided in Example 2 of the present invention was measured;

[0022] Figure 4 The standard activity of yeast A provided in Example 2 of the present invention;

[0023] Figure 5 The activity of the yeast B provided in Example 2 of the present invention was measured;

[0024] Figure 6 The standard activity of yeast B provided in Example 2 of the present invention;

[0025] Figure 7 The activity of yeast C provided in Example 2 of the present invention was measured;

[0026] Figure 8 The standard activity of yeast C provided in Example 2 of the present invention;

[0027] In the above figures, 1. liquid storage tank; 2. channel; 3. plugging agent; 4. diluted microbial suspension. DETAILED DESCRIPTION

[0028] The technical scheme in the specific embodiment of the present invention will be described in detail and completely in conjunction with the accompanying drawings. Obviously, the described embodiments are only some specific implementation methods of the overall technical scheme of the present invention, rather than all implementation methods. Based on the overall concept of the present invention, all other embodiments obtained by ordinary technicians in this field fall within the scope of protection of the present invention.

[0029] The present invention proposes a solid-state fermentation sample microorganism rapid detection kit, comprising a separation unit for separating microorganisms from a solid-state fermentation sample, a dilution unit for diluting the separated microorganism suspension with a thickener solution, and a chip for incubating and culturing the diluted microorganism suspension 4 so that the microorganism single cells grow into cell clusters and then performing identification and statistics; the separation unit comprises: a filter element, which is coated on the outside of the solid-state fermentation sample and has a plurality of filter holes; a separation element, in which a solution is provided in a cavity, and the filter element is immersed in the solution; an oscillating element, which is used to oscillate the separation element so that the microorganisms in the solid-state fermentation sample coated in the filter element flow out of the filter element from the filter holes and enter the separation element; Figure 1 As shown, the chip includes: a channel 2, in which the diluted microbial suspension 4 is incubated and cultured until the microbial single cell grows into a cell cluster, and then identification and statistics are performed, and the aspect ratio of the dimension in the channel 2 is less than 40:1; a liquid storage tank 1, connected to the channel 2, for inputting the diluted microbial suspension 4 into the channel 2, and the inner cross-sectional width of the liquid storage tank 1 is greater than the droplet width of the diluted microbial suspension 4; the amount of the diluted microbial suspension 4 added to the channel 2 is controlled to be 80-120% of the volume of the channel 2, and when the amount of the diluted microbial suspension 4 added to the channel 2 is greater than the volume of the channel 2, the area of ​​the diluted microbial suspension 4 exceeding the channel 2 on the bottom surface of the liquid storage tank 1 does not exceed 60% of the bottom surface area of ​​the liquid storage tank 1.

[0030] The solid-state fermentation sample microbial rapid detection kit uses a separation unit to achieve rapid separation of microorganisms and large particle impurities, and uses a dilution unit to dilute the microbial suspension, thereby reducing the influence of impurities with smaller particle sizes on the process of identifying cell clusters. At the same time, the structure of the chip is limited to ensure that there are no bubbles in the microbial suspension in channel 2, ensure the stability of the microbial suspension in channel 2, and realize static culture of microorganisms in chip channel 2. The budding and proliferating cells will stay and attach to the parent cell enrichment, while the cells that have not proliferated will still maintain a single cell state, so that the two can be distinguished by imaging, thereby improving the accuracy of detection.

[0031] Specifically, the above technical solution defines that the width of the inner cross section of the liquid reservoir 1 is greater than the width of the droplet of the diluted microbial suspension 4. The reason is that when the width of the liquid reservoir 1 is less than the width of the droplet, the air at the lower end of the droplet is difficult to be discharged to the outside of the liquid reservoir 1, thereby generating bubbles, which affects the siphon effect in the next step. Furthermore, the cross section of the liquid reservoir 1 is narrow at the top and wide at the bottom, which can better avoid the problem of bubbles when the diluted microbial suspension 4 is introduced from the liquid reservoir 1 into the channel 2. The width of the cross section at the narrowest point is greater than the width of the droplet of the diluted microbial suspension 4. The above technical solution also defines that the aspect ratio of the inner dimension of the channel 2 is less than 40:1. Through the aspect ratio design of the channel 2, the effect of the sample capillary into the channel 2 without bubbles is achieved, and at the same time, the imaging of the large-capacity channel 2 is also satisfied. It should be noted that if the aspect ratio of the channel 2 is greater than 40:1, the problem of insufficient surface tension of the liquid to fill the chamber will occur, thereby generating bubbles. The above technical solution also specifically limits the amount of liquid added to the diluted microbial suspension 4 in the channel 2. The reason is that, by controlling the amount of liquid added, it is ensured that the liquid fills the channel 2, the liquid level is stable, and the sample imaging requirements are met. Specifically, the amount of liquid added is controlled within ±20% of the volume of the channel 2, and the maximum does not exceed 60% of the bottom area of ​​the liquid storage tank 1. When there is too much liquid, it fills the bottom of the liquid storage tank 1 and becomes a communicating vessel, causing the liquid level to be unstable, thereby affecting the stability of the diluted microbial suspension 4 in the channel 2, affecting the subsequent detection efficiency and accuracy.

[0032] It should be noted that the present invention limits the incubation and culture of microorganisms in the chip channel 2 until the single microbial cells grow into cell clusters for identification and statistics, rather than incubating and culturing the microorganisms until the single microbial cells grow into cell clusters and then introducing them into the chip for identification and statistics. The reason is that the chip culture of the present invention is a plate culture strategy with visible process and higher detection sensitivity. The cells in the chip are in a discrete and relatively static state of single cells. The budding and proliferating cells will stay and attach to the enriched parent cells, while the cells that have not proliferated will still maintain a single cell state, so that the two can be distinguished by imaging. If the chip is introduced after culture, the budding and clustered bacteria will be broken up again, and this difference cannot be observed.

[0033] The above technical scheme also limits the use of a thickener solution to dilute the microbial suspension. The reason is that the thickener has a high viscosity. The thickener is used to limit the movement of the microorganisms in the chip, thereby improving the clarity of the imaging and the accuracy of the analysis. By adding a thickener, the Brownian motion of the microorganisms is reduced, and the stability requirements for microbial imaging are met. Furthermore, the thickener is preferably a transparent thickener that does not affect cell growth. Preferably, the thickener is xanthan gum. At room temperature, a 0.1%wt xanthan gum aqueous solution can increase the viscosity by about 100 times compared to pure water, thereby weakening the movement of microorganisms in the solution and facilitating imaging. In addition, xanthan gum is a polysaccharide produced by fermentation of bacteria (mainly Xanthomonas), which is widely used in the food, pharmaceutical and cosmetic industries, has high biosafety, and will not have a visible effect on the growth and proliferation of microorganisms. Analysis: Thickeners increase the viscosity of liquids and reduce the amplitude of the Brownian motion of microorganisms.

[0034] In a preferred embodiment, the chip further comprises: an upper layer of glass, a channel 2 structural layer, and a bottom layer of glass arranged in sequence from top to bottom; the upper layer of glass, the hollowed-out portion of the channel 2 structural layer, and the bottom layer of glass enclose the channel 2; and the liquid reservoir 1 is connected to at least one end of the channel 2 through a through hole. Further, the liquid reservoir 1 is arranged in parallel along the left and right sides of the upper layer of glass, and both ends of the channel 2 are connected to the liquid reservoirs 1 arranged at both ends through through holes.

[0035] It is understandable that, in order to ensure the flux of the detection, there are several channels 2, at least one end of the several channels 2 is connected to the liquid reservoir 1 through a through hole, and preferably both ends of the channel 2 are connected to the liquid reservoirs 1 arranged on both sides. As for the structural form of the liquid reservoir 1, as long as it is connected to the channel 2 and the inner cross-sectional width of the liquid reservoir 1 is greater than the droplet width of the diluted microbial suspension 4, it can be an integral structure (several liquid reservoir 1 chambers are integrated in one structure) or a split structure (each liquid reservoir 1 corresponds to a liquid reservoir 1 chamber), but in order to ensure the effect, one end of each channel 2 is connected to a liquid reservoir 1 chamber.

[0036] In a preferred embodiment, the chip incubates and cultivates the diluted microbial suspension 4 in the channel 2 under a constant humidity environment; or after the diluted microbial suspension 4 is injected into the channel 2, the reservoir 1 is blocked to isolate the channel 2 from the outside world. By adding a blocking agent 3 to the reservoir 1, the liquid in the channel 2 is non-volatile during the incubation process. In the above technical scheme, the constant humidity keeps the liquid non-volatile, or adds incompatible chemicals, or a barrier medium to prevent the liquid from volatilizing. Further, the blocking agent 3, the plug, or the membrane is used to block the reservoir 1 to isolate the channel 2 from the outside world. When microorganisms are cultivated in the chip, on the one hand, the amount of the suspension containing microorganisms is very small, and on the other hand, the suspension containing microorganisms has a large specific surface area in the channel 2. Therefore, the water in the suspension containing microorganisms in the channel 2 is very easy to volatilize, thereby affecting the incubation effect, and even affecting the subsequent identification detection. The limitation of the above technical scheme of the present invention avoids the evaporation of water in the sample in the chip when the microorganisms are cultivated in the chip.

[0037] In a preferred embodiment, the separated microbial suspension is diluted with a thickener solution to 1% to 99% of the original microbial suspension concentration, reducing the probability that the target microorganisms are blocked by impurities in the chip. The technical solution limits the dilution multiple of the microbial suspension with a thickener solution because the dilution multiple can ensure the detection effect while solving the interference of small particles on the detection process.

[0038] In a preferred embodiment, the filter element includes but is not limited to a filter bag, a filter membrane or a filter screen; the separation element includes but is not limited to a centrifuge tube, a test tube, and a conical flask; the solution includes but is not limited to a culture medium and a buffer. In the above-mentioned kit, the solution is used to provide the nutrients required for the growth of microorganisms; the thickener solution is used to limit the movement of microorganisms in the microbial detection chip to improve the imaging quality; the plugging agent 3 is used to prevent the evaporation of water after the sample is injected into the microbial detection chip; the filter bag, filter membrane or filter screen is used as a filter element to remove large particle impurities; the chip is used for subsequent microbial detection and analysis; the sampling spoon is used for quantitative sampling of solid-state fermentation samples. The solid-state fermentation sample microbial rapid detection kit enables the solid-state fermentation sample to be pre-treated, which is conducive to reducing costs, reducing sample losses, and improving detection efficiency.

[0039] In a preferred embodiment, a hollow structure is provided on the channel 2 structural layer between adjacent channels 2 for microscopic field focusing. By introducing a hollow structure next to the channel 2, the need for fast focusing is met. Specifically, a hollow channel 2 is designed on the upper surface of the bottom glass and the lower surface of the top glass. When focusing, the focus is directly on the upper surface of the bottom glass. The upper surface may or may not have a logo. The hollow structure has no effect on the imaging of the upper surface of the bottom glass, which is convenient for introducing logos or other optical distance measurement methods.

[0040] The present invention also provides a method for rapid detection of microorganisms in solid-state fermentation samples, which uses the solid-state fermentation sample microorganism rapid detection kit described in any of the above technical solutions to detect microorganisms in solid-state fermentation samples, including separating microorganisms from the solid-state fermentation sample through a separation unit, and diluting the microbial suspension with a thickener solution, and using a liquid storage tank 1 to input the diluted microbial suspension into a channel 2 for incubation and culture so that single microbial cells can grow into cell clusters for identification and statistics.

[0041] It should be noted that the existing chip-based microbial detection technology has some significant disadvantages when processing solid-state fermentation samples:

[0042] 1) Complex sample pretreatment: Direct injection of solid-state fermentation samples can easily lead to chip clogging, and requires tedious grinding, centrifugation and other steps, which is not only time-consuming but may also cause damage or loss of microorganisms.

[0043] 2) Severe interference from impurities: Solid particles, proteins and other macromolecular substances in solid-state fermentation samples are easily deposited in the chip, affecting the imaging quality and thus reducing the detection accuracy.

[0044] 3) High cost: Traditional pretreatment reagents and consumables are expensive, which is not conducive to large-scale application.

[0045] These disadvantages limit the wide application of the prior art in the field of solid-state fermentation sample microbial detection. The solid-state fermentation sample microbial rapid detection method of the present invention solves the above problems and has the following advantages:

[0046] 1) Simplified operation: Using filter bags, filter membranes or filter screens as filtering devices is easy to use and does not require complicated operating steps;

[0047] 2) Low cost: Filter bags, filter membranes or filter screens, thickeners and other reagents are easy to obtain and inexpensive, making them suitable for large-scale applications;

[0048] 3) Reduce sample loss: The simplified process reduces the loss of microorganisms and improves sample recovery;

[0049] 4) Improve imaging quality: The use of thickeners limits the movement of microorganisms in the chip, and the appropriate dilution ratio reduces the impact of impurities, improving the clarity of imaging and the accuracy of analysis;

[0050] 5) Shorten the detection cycle: 16 solid-state fermentation samples can be tested within 4 hours, which can be applied not only to technical research, but also to production testing, i.e. engineering;

[0051] 6) High degree of visualization: the morphology of microbial cells and cell clusters can be directly observed, and the cell type can be given in real time;

[0052] 7) The types, quantity and activity of microorganisms in the mash samples can be analyzed simultaneously.

[0053] Example 1

[0054] Identification and statistics of acetic acid bacteria in vinegar mash

[0055] Accurately grasping and understanding the changes in microorganisms during vinegar fermentation is conducive to the rational control of the aged vinegar production process. Therefore, in the vinegar brewing process, rapid detection of microorganisms in vinegar mash is a very important task.

[0056] 1 g of each of the vinegar mash samples from the 1st, 3rd, 5th, 7th, 9th, 11th, 13th, 15th, 17th, 19th and 21st days of fermentation were taken and placed in different test tubes, and 20 mL of acetic acid bacteria culture medium was added, and the mixture was shaken for 5 minutes to fully release the microorganisms to obtain a sample suspension;

[0057] Take the sample suspension, filter it through a 10 μm filter membrane to obtain a microbial suspension; mix the microbial suspension and xanthan gum evenly, add it to the chip reservoir, let the mixed solution flow into the channel, and seal the reservoir with mineral oil, and place the chip in a 30°C incubator for 3 hours;

[0058] The chip was taken out for imaging detection, and the acetic acid bacteria were identified and counted through the recognition model.

[0059] The statistical results are as follows Figure 2 As shown in the figure, the number of acetic acid bacteria increased rapidly around the 13th day of fermentation and reached its peak around the 17th day.

[0060] Example 2

[0061] Identification and activity detection of yeast species in fermented grains

[0062] During the second round of fermentation of the mash, samples were taken from the mash on the third day of pile fermentation.

[0063] Take 1 g of the fermented grains sample on the third day, put it in a filter bag, and put the filter bag in a centrifuge tube. Add 20 mL of yeast culture medium to the centrifuge tube and shake for 20 minutes to fully release the microorganisms to obtain a sample suspension.

[0064] The sample suspension was mixed evenly with xanthan gum and added to the chip reservoir. The mixed solution was allowed to flow into the channel and the reservoir was sealed with mineral oil. The chip was placed in a 30°C incubator for 3 hours.

[0065] The chip was taken out for imaging detection, and the yeast was identified through the recognition model and the activity was analyzed.

[0066] The statistical results of the activity of three yeasts A, B, and C are as follows Figure 3-8 As shown, Figure 3 , 5 , 7 are the activity data of three kinds of yeasts. Figure 4 , 6 8 are the low, medium and high standard activity data of the three yeasts used for comparison. Figure 3-8 It can be seen that the activities of the three yeasts measured are all medium.

Claims

1. A solid-state fermentation sample microbial rapid detection kit, characterized in that: It includes a separation unit for separating microorganisms from a solid-state fermentation sample, a dilution unit for diluting the separated microorganism suspension with a thickener solution, and a chip for incubating and culturing the diluted microorganism suspension to allow single microorganism cells to grow into cell clusters and then performing identification and statistics; The separation unit includes: A filter element, which is coated on the outside of the solid-state fermentation sample and has a plurality of filter holes; The separation element has a cavity with a solution, and the filter element is immersed in the solution; An oscillating member, used for oscillating the separation member so that the microorganisms in the solid-state fermentation sample coated in the filter member flow out of the filter member through the filter holes and enter the separation member; Chips include: Channel, the diluted microbial suspension is incubated and cultured in the channel until the microbial single cells grow into cell clusters, and then identification and statistics are performed. The aspect ratio of the dimensions in the channel is <40:1; A liquid storage tank connected to the channel and used to input the diluted microbial suspension into the channel, wherein the inner cross-sectional width of the liquid storage tank is greater than the droplet width of the diluted microbial suspension; The amount of the diluted microbial suspension added to the channel is controlled to be 80-120% of the channel volume, and when the amount of the diluted microbial suspension added to the channel is greater than the channel volume, the area of ​​the diluted microbial suspension exceeding the channel on the bottom of the reservoir does not exceed 60% of the bottom of the reservoir.

2. The solid-state fermentation sample microbial rapid detection kit according to claim 1, characterized in that: The chip also includes: an upper layer of glass, a channel structure layer, and a bottom layer of glass arranged in sequence from top to bottom; the upper layer of glass, the hollowed-out portion of the channel structure layer, and the bottom layer of glass form a channel; and the liquid storage tank is connected to at least one end of the channel through a through hole.

3. The solid-state fermentation sample microbial rapid detection kit according to claim 1, characterized in that: The liquid storage tanks are arranged in parallel along the left and right sides of the upper glass, and the two ends of the channel are communicated with the liquid storage tanks arranged at the two ends through through holes.

4. The solid-state fermentation sample microbial rapid detection kit according to claim 1, characterized in that: The chip incubates and cultivates the diluted microbial suspension in the channel under a constant humidity environment; or after the diluted microbial suspension is injected into the channel, the liquid storage tank is blocked to isolate the channel from the outside.

5. The solid-state fermentation sample microbial rapid detection kit according to claim 4, characterized in that: The reservoir is sealed with a plugging agent, a plug, or a membrane to isolate the channel from the outside world.

6. The solid-state fermentation sample microbial rapid detection kit according to claim 1, characterized in that: The separated microbial suspension is diluted with a thickener solution to a concentration of 1% to 99% of the original microbial suspension.

7. The solid-state fermentation sample microbial rapid detection kit according to claim 1, characterized in that: Filters include but are not limited to filter bags, filter membranes or filter screens; separation components include but are not limited to centrifuge tubes, test tubes, and conical flasks; solutions include but are not limited to culture media and buffer solutions.

8. The solid-state fermentation sample microbial rapid detection kit according to claim 1, characterized in that: A hollow structure is provided on the channel structure layer between adjacent channels for focusing the microscopic field of view.

9. A method for rapid detection of microorganisms in solid-state fermentation samples, characterized in that: The solid-state fermentation sample microorganism rapid detection kit according to any one of claims 1 to 8 is used to detect the solid-state fermentation sample microorganism.

10. The method for rapid detection of microorganisms in solid-state fermentation samples according to claim 9, characterized in that: include: The solid-state fermentation sample is separated into microorganisms by a separation unit, and the microbial suspension is diluted with a thickener solution. The diluted microbial suspension is input into the channel through a storage tank for incubation and culture so that the microbial single cells can grow into cell clusters for identification and statistics.

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