CN103063633A - System capable of automatically detecting bacteria in water - Google Patents

Abstract

The invention relates to a system capable of automatically detecting bacteria in water. The system capable of automatically detecting the bacteria in the water comprises a sampling system, a filter system, a rotary table and a detection and analysis system. Filter membrane is arranged for the system to perform bacteria gathering, and the effect of efficient and quick separation can be achieved, and specific fluorochrome is further combined to enable the detection of the bacteria to have specificity. The system capable of automatically detecting the bacteria in the water is convenient and fast to use, and has high application value.

Description

A system for rapid and automatic detection of bacteria in water

technical field

The invention belongs to the field of analysis and detection equipment, and in particular relates to a water bacteria detection system.

Background technique

Bacteria detection systems are commonly used equipment in laboratories, food processing plants and drinking water treatment plants. It is an essential tool to ensure the quality of food and drinking water and human health. Although the existing methods for detecting bacteria in water have their own advantages, most of them have their unavoidable shortcomings. For example, the sample pretreatment and detection process of many instruments is time-consuming, and the bacteria content cannot be monitored in time to do preventive work, thus harming the human body. Health is at stake. Another example is that some detection systems and instruments are large in size, inconvenient to carry and place, and are not suitable for promotion and use at the grassroots level.

Contents of the invention

The invention provides an easy-to-operate, fast and sensitive water bacteria detection system. Specifically, the following technical solutions are included:

A system for quickly and automatically detecting bacteria in water, comprising a sampling system, a filtration system, a turntable, and a detection and analysis system; the sampling system includes a pure water tank, a sample tube, a pure water valve for controlling the flow rate of pure water, and a Sample valve, mixing tube for mixing samples and pure water; the pure water tank is connected to the mixing tube with a pure water valve in between; the sample tube is connected to the mixing tube with a sample valve in between; the mixing tube is connected to the filtration system pipeline; The filtration system includes a spray head connected to the mixing tube of the sampling system, a filter head with a filter membrane directly below the spray head, a vacuum chamber directly below the filter head, and a vacuum pump connected to the vacuum chamber; the spray head, filter head, and vacuum chamber are closely sized Cooperate, turn on the vacuum pump, suck the liquid in the nozzle into the filter head, filter it through the filter membrane, and enter the vacuum chamber after filtering; between; the detection and analysis system sequentially includes a light excitation unit, a condenser, an eyepiece, a charge-coupled device (CCD) detection device, and a data analysis display unit; the filter head can be rotated into between the condenser and the eyepiece through a turntable. The light emitted by the light excitation unit hits the filter membrane of the filter head through the condenser lens, and the fluorescent dye combined with the bacteria on the excitation filter membrane emits fluorescence. The fluorescence is converted into a data signal by the CCD detection device through the eyepiece, and then transmitted to the data analysis. Display unit.

In order to reduce the dead angle of materials and bacteria, the pipes used in the device can be made of smooth stainless steel, which is corrosion-resistant and can reduce the probability of materials, bacteria and pipe walls adhering.

In order to achieve the purpose of continuous operation, a plurality of filter heads, preferably 5, are connected on the turntable. When in use, it can test 5 samples at the same time, which is very convenient. The spray head and the filter head, and the filter head and the vacuum chamber can be connected by threads.

There is a lifting table below the vacuum chamber, which can support and fix the vacuum chamber and make the vacuum chamber rise and fall together. By controlling the tightness between the vacuum chamber and the filter head, the airtightness of the filtration system can be controlled. Certainly, the connection mode between the vacuum chamber and the filter head can also be threaded or the like.

The bottom of the vacuum chamber is connected with the waste liquid bottle. A valve is arranged between the waste liquid bottle and the vacuum chamber, and the waste liquid in the vacuum chamber can be stored in the waste liquid bottle.

Sealing rings are provided at the junction of the spray head and the filter head, and the junction of the filter head and the vacuum chamber.

The filter head is equipped with a disposable filter membrane with a pore diameter of 0.22 μm. In this way, after the filter membrane is used, it is very convenient to directly discard and replace the new filter membrane.

The data display system can also be used to set parameters and display data, and set different detection parameters according to the emission wavelengths of different fluorescent dyes to detect samples.

The beneficial effect of the invention is that it is very convenient and can continuously detect bacteria in water samples, especially pathogenic bacteria. Because the system is equipped with a filter membrane for bacterial enrichment, it can achieve high-efficiency and rapid separation effects. Combined with specific fluorescent dyes, the detection of bacteria can be specific.         

Description of drawings

The present invention will be further described in detail below in conjunction with the accompanying drawings and the implementation method of the present invention.

Figure 1 The system of the present invention for rapid and automatic detection of bacteria in water.

1 sample tube, 2 pure water tank, 3 sample valve, 4 pure water valve, 5 mixing tube, 6 nozzle, 7 filter head, 8 vacuum chamber, 9 vacuum pump, 10 turntable, 11 light excitation unit, 12 condenser, 13 eyepiece, 14 CCD detection device, 15 data analysis display unit, 16 waste liquid bottle, 17 lifting platform.

Fig. 2 The detection results of E.coli O157:H7 in drinking water using the system of the present invention.

Detailed ways

In order to make the present invention clearer, the present invention will be further described in detail in combination with examples. The specific embodiments described here are only used to explain the present invention, not to limit the present invention.

Embodiment 1 The system of the present invention

The system for rapid and automatic detection of bacteria in water shown in Figure 1 includes a sampling system, a filtration system, a turntable, and a detection and analysis system; the sampling system includes a sample tube 1, a pure water tank 2, and a pure water valve 4 for controlling the flow rate of pure water , the sample valve 3 for controlling the flow rate of the sample, the mixing tube 5 for mixing the sample and pure water; The water valve 4; the mixing pipe 5 is connected to the filtration system through pipelines; the filtration system includes a sampling nozzle 6 connected to the mixing pipe 5 of the sampling system, a filter head 7 loaded with a filter membrane just below the nozzle 6, and a filter head 7 directly below the nozzle 6. The vacuum chamber 8 below, the vacuum pump 9 that is connected with the vacuum chamber 8; The size of the nozzle 6, the filter head 7, and the vacuum chamber 8 are adapted to each other, and they are connected by threads, and the liquid (sample liquid) in the nozzle 6 is opened by opening the vacuum pump 9. , pure water, or mixed solution, cleaning solution, etc.) are sucked into the filter head 7 to filter, and after filtering, enter the vacuum chamber, and the bacteria are trapped on the filter membrane loaded by the filter head 7; the turntable 10 is connected with the filter head 7, and passes through Rotate the turntable 11 to spin the filter head 7 away or screw it in between the nozzle 6 of the filter system and the vacuum chamber 8; the detection and analysis system sequentially includes a light excitation unit 11, a condenser lens 12, an eyepiece 13, a CCD detection device 14, and a data analysis display unit 15; the filter head 7 is screwed into between the condenser lens 12 and the eyepiece 13 through the turntable 10, and the light emitted by the light excitation unit 11 hits the filter sheet of the filter head 7 through the condenser lens 12 to excite the fluorescence combined with bacteria on the filter sheet The dye emits fluorescence, which is converted into a data signal by the CCD detection device 14 through the eyepiece 13 and then transmitted to the data analysis and display unit 15 . Of course, a lifting platform 17 can also be arranged below the vacuum chamber 8 to support the vacuum chamber to rise and fall, and can also prevent the vacuum chamber from falling directly.

When in use, the sample tube 1 and the pure water tank 2 respectively control the switch and flow rate through the pure water valve 3 and the sample valve 4, and the sample and pure water are mixed in the mixing tube 5 and sent to the nozzle 6 by starting the vacuum pump 9, and sprayed on the On the filter membrane of the filter head 7, so as to achieve the effect of quickly and effectively enriching the bacteria in the sample, the filtered waste liquid enters the vacuum chamber 8, and is discharged to the waste liquid bottle 16 through the valve 18 control. When the sample in the sample tube 1 is blotted dry, the sample valve 3 is closed, and the pure water can wash down the bacteria remaining in the tube. When testing the next sample, close the pure water valve 4, open the sample valve 3, decompress the vacuum system, unscrew the interface between the vacuum chamber 8 and the filter head 7, automatically rotate the filter head 7 to the condenser lens 12, and pass through the light excitation unit 11, the fluorescent dye combined with the bacteria is excited to emit fluorescence. After the fluorescence enters the eyepiece 13, the fluorescent signal can be converted into a data signal by the CCD detection device 14, and then sent to the data analysis display unit 15.

Before the sample detection, the data analysis display unit can perform multi-parameter settings according to different fluorescence emission wavelengths, so that multiple bacteria can be detected in the same sample.

Example 2 A system for rapid and automatic detection of bacteria in water according to the present invention is used for the detection of E.coli O157:H7 in drinking water

1) Selection of fluorescent dyes: Most of the commonly used fluorescent dyes are fluorescent substances found in natural algae, such as PE, FITC and APC, etc. These fluorescent dyes are easy to quench and have no specificity. Here we can use Quantum Dots (QDs). QDs are an inorganic nanomaterial with fluorescence. Compared with other organic dyes mentioned above, QDs have stronger fluorescence intensity and no quenching. More importantly, the modified QDs can be combined with antibodies, proteins, etc., which makes the detection of bacteria specific.

2) Pretreatment of QDs markers: use the prepared anti -E.coli O157:H7 polyclonal antibody or monoclonal antibody to couple with QDs, and put the coupled complex in a 4°C refrigerator for use in subsequent experiments .

3) Use the system for detection: take 10 mL of the sample to be tested into the injection tube (1), rotate the filter head carrying the blank filter membrane to the top of the vacuum chamber through the turntable, and tighten the threaded interface between the vacuum chamber and the filter head , so that it is in a closed state. Turn on the vacuum pump (9), the sample valve, and the pure water valve, so that the sample and pure water are mixed in the mixing tube and sprayed on the filter membrane through the nozzle. When the sample is blotted dry, close the injection valve, at this time, the sterile water will flush the residual bacteria in the pipeline. After a quick flush for about 10 seconds, close the pure water valve, open the injection valve, and connect the air to decompress. Unscrew the thread of the vacuum chamber and the filter head, loosen the filter head, and drop the above-mentioned E.coli O157:H7 antibody marked with QDs on the filter membrane that has collected the bacteria. If there is E.coli O157 in the sample to be tested :H7, the antigen on its surface will react with the labeled antibody, so that E.coli O157:H7 is indirectly linked to QDs. After dyeing, rotate the filter head under the condenser for detection, and the detection results are shown in Figure 2. The detection result is analyzed by the system and displayed by the data analysis display unit. While the system is performing detection and analysis, another sample can continue to be injected and filtered according to the above steps to prepare for the next detection and analysis.

Example 3 The detection of the total number of bacteria in drinking water

Collect 10 mL of the water sample to be tested and add it to the sampling tube, start the sample injection and filtration as described in Example 1, after the filtration is completed, add FITC dye dropwise on the filter membrane to stain the bacteria, add 1 μl FITC dye dropwise to each ml sample, and incubate for 30 min. After staining, rotate the filter head to the condenser for detection. The detection result will be displayed by the data analysis display unit.

Claims (6)

1. A system for rapid and automatic detection of bacteria in water, comprising a sampling system, a filter system, a turntable, and a detection and analysis system; the sampling system includes a pure water tank, a sample tube, a pure water valve for controlling the flow rate of pure water, and a control sample Flow rate sample valve, mixed sample and pure water mixing tube; pure water tank is connected to the mixing tube with a pure water valve in between; the sample tube is connected to the mixing tube with a sample valve in between; the mixing tube is connected to the filtration system pipeline; The filtration system includes a spray head connected to the mixing tube, a filter head with a filter membrane directly below the spray head, a vacuum chamber directly below the filter head, and a vacuum pump connected to the vacuum chamber; the size of the spray head, the filter head, and the vacuum chamber are closely matched , turn on the vacuum pump, suck the liquid in the nozzle into the filter head, filter it through the filter membrane, and then enter the vacuum chamber; Between; The detection and analysis system includes an optical excitation unit, a condenser, an eyepiece, a CCD detection device, and a data analysis display unit in turn; the filter head is rotated into between the condenser and the eyepiece through a turntable, and the light emitted by the optical excitation unit passes through the condenser to the filter On the filter membrane of the head, the fluorescent dye combined with the bacteria on the excitation filter membrane emits fluorescence, and the fluorescence is converted into a data signal by the CCD detection device through the eyepiece, and then transmitted to the data analysis display unit. 2. The system according to claim 1, characterized in that a plurality of filter heads, preferably 5, are connected to the turntable. 3. The system according to claim 1, characterized in that a lifting platform is provided under the vacuum chamber. 4. The system of claim 1, wherein the bottom of the vacuum chamber is connected to a waste liquid bottle. 5 . The system according to claim 1 , wherein the filter head is loaded with a disposable filter membrane with a pore diameter of 0.22 μm. 6. The system according to claim 1, characterized in that there are sealing rings at the junction of the spray head and the filter head, and the junction of the filter head and the vacuum chamber.

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