Microorganism sampling and detecting device for plateau honey

Abstract

The invention provides a plateau honey microorganism sampling and detecting device, and belongs to the technical field of honey production devices.The plateau honey microorganism sampling and detecting device comprises an operation table, an operation ring, a sampling vessel, a connecting column, a sampling tube, a reaction tube and an ATP fluorescence detector, a base is installed on the operation table, a supporting column is installed on the base, and the operation ring is installed at the top end of the supporting column; a motor group is uniformly mounted on the top wall of the operating ring, a sampling vessel covers the top wall of the operating ring, a connecting column is connected with a motor power output end of the motor group, a sampling tube detachably sleeves the lower end of the connecting column, a reaction tube detachably sleeves the lower part of the sampling tube, and the ATP fluorescence detector is mounted on one side of the operating table. By means of the plateau honey microorganism sampling detection device, the problems that when a sampling detection device used by a traditional plate culture method or a PCR technology is applied to the plateau honey microorganism sampling detection process, representative samples are difficult to obtain efficiently, and the detection time is long can be solved.

Description

Technical Field

[0001] The present invention belongs to the technical field of honey production devices, and particularly relates to a microbial sampling and detection device for plateau honey. Background Art

[0002] Plateau honey is produced in high-altitude areas, usually with an altitude of ≥3000 meters. The unique natural environment endows honey with special nutritional components. The content of antioxidant active substances such as polyphenols and flavonoids is 20% - 30% higher than that of ordinary honey. During the transportation from plateau to plain, due to the change in air pressure, the residual air in the sealed container expands, which will cause the honey to undergo phase separation. The honey shows liquid-solid stratification, with the upper layer being thin and liquid and the lower layer being thick. With the temperature change during transportation or poor container sterilization conditions, the water in the thin layer will ferment and produce microbial contamination. At the same time, temperature fluctuations will cause the honey to melt and crystallize repeatedly, destroying the natural antibacterial barrier of the honey and contaminating the thick honey with microorganisms.

[0003] After the honey is contaminated by microorganisms, it will not only cause the honey to ferment and produce peculiar smells, but also make it carry foodborne pathogens such as Salmonella. However, after the plateau honey is transported to the plain area, it needs to pass through customs quickly and enter the cold chain storage. If the microbial detection is not completed in time, the following risks may occur: First, the contaminated honey can exponentially proliferate microorganisms within one day at room temperature, destroying the nutritional components, quality and safety of the honey. Second, the traditional detection cycle is as long as 2 - 3 days, resulting in the honey staying in a non-temperature-controlled environment, accelerating phase separation and secondary contamination.

[0004] Currently, the microbial detection of plateau honey mainly relies on the plate culture method or conventional PCR technology. For example, the patent technology CN201510244394.X discloses a method for detecting microorganisms in honey. The sealed sample card is placed in an incubator for constant temperature culture at a temperature of 32 - 34°C for 36 hours. Observe whether bubbles are generated on the water surface of the sample card. If bubbles are generated, it proves the presence of microorganisms; if no bubbles are generated, it proves the absence of microorganisms. Its detection time is more than 36 hours, and it is difficult to be applicable to the honey that needs to be stored urgently after being transported from the plateau area. The detection time of the conventional PCR technology is about 4 - 6 hours.

[0005] Due to the high viscosity characteristics of honey, the existence and distribution of microorganisms in the honey that has melted repeatedly during transportation are not stable. It is difficult for the above-mentioned plate culture method or PCR technology to efficiently obtain representative samples by using cotton swab wiping or liquid aspiration methods. It is necessary to repeat multiple steps such as sampling, dilution, and mixing, which further prolongs the detection time. The PCR detection is extended to about one day, and it is easy to introduce operation contamination. Therefore, the present application proposes a microbial sampling and detection device for plateau honey. Summary of the Invention

[0006] To overcome the problems in the background art, the present invention has developed a microbial sampling and detection device for plateau honey, which solves the problems of difficult to efficiently obtain representative samples and long detection time in the process of applying the sampling and detection devices used in traditional plate culture method or PCR technology to the microbial sampling and detection of plateau honey.

[0007] To achieve the above object, the present invention is realized through the following technical solutions: A microbial sampling and detection device for plateau honey, comprising an operation table, an operation ring, a sampling dish, a connecting column, a sampling tube, a reaction tube and an ATP fluorescence detector. A base is installed on the operation table, a support column is installed on the base, a central shaft rod is installed at the center of the operation ring, the lower end of the central shaft rod is rotatably installed in the jack opened at the top end of the support column, and a motor group is evenly installed on the top wall of the operation ring. The sampling dish covers the top wall of the operation ring. The connecting columns are evenly arranged below the operation ring and the upper ends pass through the through holes opened on the operation ring and are connected to the motor power output ends of the motor group. The sampling tube is detachably sleeved on the lower end of the connecting column, and the reaction tube is detachably sleeved on the lower part of the sampling tube. The ATP fluorescence detector is installed on one side of the operation table, and the fluorescence detection slot of the ATP fluorescence detector is correspondingly arranged below the connecting column.

[0008] Further, a first limiting platform is arranged on the lower part of the connecting column, and a convex ring is arranged on the connecting column below the first limiting platform.

[0009] Further, the tube bodies of the sampling tube and the reaction tube are made of disposable transparent material.

[0010] Further, the upper part of the sampling tube is a sleeve, the bottom end of the sleeve is connected with a sampling column, a second limiting platform is arranged on the outer side of the sleeve, and a convex ring is arranged on the outer wall of the sleeve below the second limiting platform.

[0011] Further, the reaction tube is a columnar test tube, the upper part of the reaction tube is sleeved on the outer side of the sampling tube, and when the tube orifice of the reaction tube abuts against the second limiting platform, the sampling column extends into the middle and lower part of the reaction tube.

[0012] Further, a detection platform is arranged on the ATP fluorescence detector, the fluorescence detection slot correspondingly runs through the detection platform in an arc shape along the rotation track of the connecting column, and a mask is slidably installed on the outer wall of the detection platform on one side of the fluorescence detection slot.

[0013] Further, a first motor is installed in the operation table, the power output end of the first motor is rotatably connected with the lower end of the support column, a limiting pin is arranged at the lower end of the central shaft rod and fixed with the jack, the mask is installed on the detection platform through an electric slide rail, and the PLC controller installed in the ATP fluorescence detector is respectively connected with the first motor and the electric slide rail.

[0014] A microbial sampling and detection method for plateau honey, comprising the following steps: 1) Sample preparation: Place a release paper on the sampling dish and spread a certain amount of honey to be tested evenly on the release paper; 2) Honey sampling: Put the sampling tube on the lower end of the connecting column and make it contact with the limit table. Remove the operating ring from the supporting column. Hold the handle in the middle of the operating ring and insert the sampling column into the sampling dish. Gently rotate it to evenly pick up the honey on the release paper. 3) Honey dilution: Add sterile diluent to the reaction tube, put the reaction tube on the lower end of the sampling tube and make it contact with the second limit table, insert the sampling column into the sterile diluent, install the operating ring back to the connecting column, start the motor group and mix for 10 minutes until it is completely dissolved.

[0015] 4) ATP extraction: Remove the diluted reaction tube, add ATP lysis reagent and fix it to the original sampling tube, and start the motor group to lysis and mix.

[0016] 5) Bioluminescence Detection: Remove the reaction tube after lysis, add luciferase reagent and fix it to the original sample tube, start the motor to mix, turn the operating ring to move the reaction tube into the fluorescence detection tank, and the ATP fluorescence detector automatically records the relative light unit (RLU) value; 6) Data analysis: Convert the RLU value to the number of microorganisms to determine whether the sample meets safety standards.

[0017] Beneficial effects of the present invention: In view of the high viscosity of honey, this application designs an operating ring, a sampling dish, a sampling tube, and a reaction tube. By cooperating with the sampling dish, the operating ring and the sampling tube, representative samples can be efficiently obtained. At the same time, the structural design of the operating ring, the sampling tube, and the reaction tube, combined with the ATP fluorescence detector, can make the detection steps more efficient and shorten the detection time, solving the problem that the sampling and detection devices used in traditional plate culture methods or PCR technologies are difficult to obtain representative samples efficiently and the detection time is long when applied to the sampling and detection process of plateau honey microorganisms. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 It is the overall structural diagram of the present invention; Figure 2 It is a storage schematic diagram of the present invention; Figure 3 It is a structural diagram of the operation ring of the present invention; Figure 4 This is a diagram of the connecting column installation structure of the present invention; Figure 5 This is a structural diagram of the connecting column, sampling tube, and reaction tube of the present invention; Figure 6 It is a reagent reaction installation structure diagram of the present invention; Figure 7 It is a structural diagram of the operating table of the present invention; Figure 8 It is a structural diagram of the ATP fluorescence detector of the present invention.

[0019] 1-operating table, 11-base, 12-support column, 2-operating ring, 21-central axis, 22-motor unit, 3-sampling dish, 4-connecting column, 41-limiting table one, 5-sampling tube, 51-casing, 52-sampling column, 53-limiting table two, 6-reaction tube, 7-ATP fluorescence detector, 71-fluorescence detection tank, 72-detection table, 73-shield. DETAILED DESCRIPTION

[0020] In order to make the objectives, technical solutions and beneficial effects of the present invention more clear, preferred embodiments of the present invention will be described in detail below with reference to the accompanying drawings to facilitate understanding by technicians.

[0021] See Figure 1-8 , a plateau honey microorganism sampling and detection device includes an operating table 1, an operating ring 2, a sampling dish 3, a connecting column 4, a sampling tube 5, a reaction tube 6 and an ATP fluorescence detector 7. The operating table 1 is equipped with a base 11, a supporting column 12 is installed on the base 11, a central axis rod 21 is installed at the center of the operating ring 2, and the lower end of the central axis rod 21 is rotatably installed in the socket opened at the top of the supporting column 12. A motor group 22 is evenly installed on the top wall of the operating ring 2, and the sampling dish 3 is covered on the top wall of the operating ring 2. The connecting column 4 is evenly arranged below the operating ring 2 and the upper end passes through the through hole opened on the operating ring 2 and is connected to the motor power output end of the motor group 22. The sampling tube 5 is detachably sleeved on the lower end of the connecting column 4, and the reaction tube 6 is detachably sleeved on the lower part of the sampling tube 5. The ATP fluorescence detector 7 is installed on one side of the operating table 1, and the fluorescence detection slot 71 of the ATP fluorescence detector 7 is correspondingly arranged below the connecting column 4.

[0022] During use, the sampling tube 5 can be sleeved on the lower end of the connecting column 4 and multiple ones can be evenly installed. The operating ring 2 can be removed from the supporting column 12. The diameter of the sampling dish 3 is larger than the spacing between a group of relative connecting columns 4 but smaller than the diameter of the operating ring 2. The honey sample can be evenly spread in the sampling dish 3. The sampling tube 5 installed on the operating ring 2 can simultaneously sample multiple points in the sampling dish 3, thereby improving the sampling efficiency and efficiently obtaining representative samples. The sampling area is evenly covered to ensure comprehensive detection and avoid omissions. Due to the high viscosity of honey, the honey sample dipped after the sampling tube 5 is gently rotated can adhere to the end of the sampling tube 5. At this time, the reaction tube 6 with the reagent added in advance can be connected to the sampling tube 5. By starting the motor group 22, the connecting column 4 is rotated to mix the honey and the reagent. The sample in the reaction tube 6 that has finally completed the reaction can be inserted into the fluorescence detection slot 71 for detection by moving the operating ring 2. The whole process is convenient and fast, and representative samples can be obtained efficiently while shortening the detection time, so that the plateau honey that has been transported over long distances can be stored in the shortest time.

[0023] Refer to Figure 5-6 , a first limiting platform 41 is provided at the lower part of the connecting column 4, a convex ring is provided on the connecting column 4 below the first limiting platform 41, the first limiting platform 41 can ensure that the sampling tube 5 is tightly installed with the connecting column 4 and the installation positions of multiple sampling tubes 5 are kept consistent, and the convex ring can increase the firmness and sealing performance of the installation of the sampling tube 5.

[0024] Refer to Figure 5-6 , the tube bodies of the sampling tube 5 and the reaction tube 6 are made of disposable transparent material tubes, which can avoid bacterial contamination and facilitate the addition of reagents to observe the reaction.

[0025] Refer to Figure 5-6 , the upper part of the sampling tube 5 is a sleeve 51, the bottom end of the sleeve 51 is connected with a sampling column 52, a second limiting platform 53 is arranged on the outer side of the sleeve 51, and a convex ring is arranged on the outer wall of the sleeve 51 below the second limiting platform 53. The sleeve 51 can be fixed with the connecting column 4, which is convenient for replacement after use and solves the problem of honey cleaning. The sampling column 52 is convenient for dipping and attaching honey samples. The second limiting platform 53 can keep the installation positions of multiple reaction tubes 6 consistent, and the convex ring can increase the firmness and sealing performance of the installation of the reaction tube 6.

[0026] Refer to Figure 5-6 , the reaction tube 6 is a cylindrical test tube, the upper part of the reaction tube 6 is sleeved on the outer side of the sampling tube 5, and when the tube orifice of the reaction tube 6 abuts against the second limiting platform 53, the sampling column 52 extends into the reagent in the middle and lower part of the reaction tube 6 to allow the honey sample to fully contact with the reagent.

[0027] Refer to Figure 7-8 , a detection platform 72 is arranged on the ATP fluorescence detector 7, a fluorescence detection groove 71 corresponding to the rotation track of the connecting column 4 is arc-shaped and penetrates through the detection platform 72, and a mask 73 is slidably installed on the outer wall of the detection platform 72 on one side of the fluorescence detection groove 71. The arc-shaped fluorescence detection groove 71 is convenient for directly rotating the operation ring 2 of the sample after the reaction is completed into the detection area of the fluorescence detection groove 71 and removing it from the detection area after detection, so as to improve the detection efficiency and operation convenience and shorten the detection time. The mask 73 can slide back and forth to shield the detection area during detection.

[0028] Refer to Figure 7-8, a first motor is installed in the operating table 1. The power output end of the first motor is rotationally connected to the lower end of the support column 12. A limit pin at the lower end of the central shaft rod 21 is fixed to a jack. The mask 73 is installed on the detection table 72 through an electric slide rail. The PLC controller installed in the ATP fluorescence detector 7 is respectively connected to the first motor and the electric slide rail. The PLC controller can receive the feedback signals of starting detection and completing detection fed back by the detection area sensor. After starting the detection, the PLC controller can send signals to the first motor and the electric slide rail. The electric slide rail moves backward to open the mask 73. The first motor drives the support column 12 to rotate to make the reaction tube 6 enter the detection area of the fluorescence detection tank 71. After the threshold value of the distance sensor or contact sensor in the detection area changes, a signal is fed back to the PLC controller. The PLC controller sends a signal to the electric slide rail to close the mask 73. A signal of the electric slide rail or the light sensor in the detection area is fed back to the PLC controller. The PLC controller sends a signal to the fluorescence detection device in the detection area to capture and detect the luminescence value. After the data acquisition is completed, the fluorescence detection device feeds back a signal to the PLC controller. The PLC controller feeds back a signal to the first motor and the electric slide rail to open the mask 73 and sequentially perform sample detection in the above order. The whole process eliminates the process of manually replacing samples, improves the detection efficiency and shortens the detection time.

[0029] On the other hand, referring to Figure 1-8 , the present invention also provides a method for sampling and detecting microorganisms in plateau honey, which includes the following steps: 1) Sample preparation: Release paper is placed in the sampling dish 3, and a certain amount of honey to be detected is evenly spread on the release paper. Since the viscosity of honey is relatively high, using release paper to spread honey can avoid cleaning problems, and different samples can be replaced by replacing the release paper; 2) Honey sampling: The sampling tube 5 is sleeved on the lower end of the connecting column 4 and abuts against the first limiting platform 41. The operating ring 2 is removed from the support column 12. Hold the handle in the middle of the operating ring 2 and gently rotate the sampling column 52 into the sampling dish 3 to evenly dip the honey on the release paper. The design of the operating ring 2 enables multiple sampling columns 52 to simultaneously and evenly sample the samples in the supporting sampling dish 3, and efficiently obtain a comprehensive and representative sample; 3) Honey dilution: Sterile diluent is added to the reaction tube 6. The reaction tube 6 is sleeved on the lower end of the sampling tube 5 and abuts against the second limiting platform 53. The sampling column 52 extends into the sterile diluent. The operating ring 2 is installed back on the connecting column 4, and the motor group 22 is started to mix for 5 minutes until completely dissolved.

[0030] 4) ATP extraction: Remove the reaction tube 6 after dilution is completed. If there is a small amount of diluted sample, directly add the ATP lysis reagent and then fix it with the original sampling tube 5. If there is a large amount of diluted sample, take a certain amount of the sample, add it to a new reaction tube 6, add the ATP lysis reagent, and then fix it with the original sampling tube 5. Start the motor group 22 to lyse and mix evenly.

[0031] 5) Bioluminescence detection: Remove the reaction tube 6 after lysis is completed. If there is a small amount of lysed sample, directly add the luciferase reagent and then fix it with the original sampling tube 5. If there is a large amount of lysed sample, take a certain amount of the sample, add it to a new reaction tube 6, add the luciferase reagent, and then fix it with the original sampling tube 5. Start the motor group 22 to mix evenly, rotate the operation ring 2 to move the reaction tube 6 into the fluorescence detection slot 71, and the ATP fluorescence detector 7 automatically records the relative light unit RLU value; 6) Data analysis: Convert the RLU value into the number of microorganisms and determine whether the sample meets the safety standard.

[0032] Finally, it should be noted that the above preferred embodiments are only used to illustrate the technical solutions of the present invention rather than to limit them. Although the present invention has been described in detail through the above preferred embodiments, those skilled in the art should understand that various changes can be made in terms of form and details without departing from the scope defined by the claims of the present invention.

Claims

1. A microbial sampling and detection device for plateau honey, characterized in that: It includes an operating table (1), an operating ring (2), a sampling dish (3), a connecting column (4), a sampling tube (5), a reaction tube (6) and an ATP fluorescence detector (7). A base (11) is installed on the operating table (1), and a support column (12) is installed on the base (11). A central shaft rod (21) is installed at the center of the operating ring (2). The lower end of the central shaft rod (21) is rotatably installed in a jack opened at the top end of the support column (12). A motor group (22) is evenly installed on the top wall of the operating ring (2). The sampling dish (3) covers the top wall of the operating ring (2). The connecting columns (4) are evenly arranged below the operating ring (2), and the upper ends pass through through holes opened on the operating ring (2) and are connected to the motor power output ends of the motor group (22). The sampling tube (5) is detachably sleeved on the lower end of the connecting column (4), and the reaction tube (6) is detachably sleeved on the lower part of the sampling tube (5). The ATP fluorescence detector (7) is installed on one side of the operating table (1), and the fluorescence detection groove (71) of the ATP fluorescence detector (7) is correspondingly arranged below the connecting column (4).

2. The high-altitude honey microbial sampling and detection device according to claim 1, characterized in that: A first limiting platform (41) is arranged at the lower part of the connecting column (4), and a convex ring is arranged on the connecting column (4) below the first limiting platform (41).

3. The high-altitude honey microorganism sampling and detection device according to claim 1, wherein: The tube bodies of the sampling tube (5) and the reaction tube (6) are made of disposable transparent material tubes.

4. The high-altitude honey microbial sampling and detection device according to claim 1, wherein: The upper part of the sampling tube (5) is a sleeve (51). The bottom end of the sleeve (51) is connected with a sampling column (52). A second limiting platform (53) is arranged on the outer side of the sleeve (51), and a convex ring is arranged on the outer wall of the sleeve (51) below the second limiting platform (53).

5. The plateau honey microorganism sampling and detection device according to claim 4, wherein: The reaction tube (6) is a cylindrical test tube. The upper part of the reaction tube (6) is sleeved on the outer side of the sampling tube (5). When the tube orifice of the reaction tube (6) abuts against the second limiting platform (53), the sampling column (52) extends into the middle and lower part of the reaction tube (6).

6. The microbial sampling and detection device for plateau honey according to claim 1, wherein: A detection platform (72) is arranged on the ATP fluorescence detector (7). The fluorescence detection groove (71) corresponding to the connecting column (4) is arc-shaped and penetrates through the detection platform (72). A mask (73) is slidably installed on the outer wall of the detection platform (72) on one side of the fluorescence detection groove (71).

7. The high-altitude honey microorganism sampling and detection device according to claim 6, characterized in that: A first motor is installed in the operating table (1). The power output end of the first motor is rotatably connected with the lower end of the support column (12). A limiting pin is arranged at the lower end of the central shaft rod (21) and is fixed to the jack. The mask (73) is installed on the detection platform (72) through an electric slide rail. The PLC controller installed in the ATP fluorescence detector (7) is respectively connected with the first motor and the electric slide rail.

8. The method for microbial sampling and detection of plateau honey described in any one of claims 1 to 7, characterized in that, It includes the following steps: 1) Sample preparation: Place release paper in the sampling dish (3), and take a certain amount of honey to be detected and evenly spread it on the release paper; 2) Honey sampling: Sleeve the sampling tube (5) on the lower end of the connecting column (4) and abut against the first limiting platform (41). Remove the operating ring (2) from the support column (12), hold the handle in the middle of the operating ring (2) and extend the sampling column (52) into the sampling dish (3) and gently rotate it to evenly dip the honey on the release paper; 3) Honey dilution: Add sterile diluent into the reaction tube (6), sleeve the reaction tube (6) at the lower end of the sampling tube (5) and make it contact with the second limiting platform (53), insert the sampling column (52) into the sterile diluent, install the operation loop (2) back onto the connecting column (4), and start the motor set (22) to mix for 5 minutes until completely dissolved; 4) ATP extraction: Remove the reaction tube (6) after dilution is completed, add ATP lysis reagent and then fix it to the original sampling tube (5), and start the motor set (22) to lyse and mix evenly; 5) Bioluminescence detection: Remove the reaction tube (6) after lysis is completed, add luciferase reagent and then fix it to the original sampling tube (5), start the motor set (22) to mix evenly, rotate the operation loop (2) to move the reaction tube (6) into the fluorescence detection slot (71), and the ATP fluorescence detector (7) automatically records the relative light unit (RLU) value; 6) Data analysis: Convert the RLU value into the number of microorganisms and determine whether the sample meets the safety standard.

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