CN116840493B - Detection device and detection method for cleaning residual protein - Google Patents
Detection device and detection method for cleaning residual protein Download PDFInfo
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- CN116840493B CN116840493B CN202311109609.8A CN202311109609A CN116840493B CN 116840493 B CN116840493 B CN 116840493B CN 202311109609 A CN202311109609 A CN 202311109609A CN 116840493 B CN116840493 B CN 116840493B
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- 238000001514 detection method Methods 0.000 title claims abstract description 196
- 102000004169 proteins and genes Human genes 0.000 title claims abstract description 87
- 108090000623 proteins and genes Proteins 0.000 title claims abstract description 87
- 238000004140 cleaning Methods 0.000 title claims abstract description 45
- 239000003153 chemical reaction reagent Substances 0.000 claims abstract description 254
- 238000005070 sampling Methods 0.000 claims abstract description 134
- 238000003860 storage Methods 0.000 claims abstract description 126
- 238000007789 sealing Methods 0.000 claims abstract description 80
- 238000012360 testing method Methods 0.000 claims abstract description 24
- 238000003780 insertion Methods 0.000 claims abstract description 17
- 230000037431 insertion Effects 0.000 claims abstract description 17
- 239000012295 chemical reaction liquid Substances 0.000 claims description 42
- 238000006243 chemical reaction Methods 0.000 claims description 28
- 230000003287 optical effect Effects 0.000 claims description 28
- 238000007790 scraping Methods 0.000 claims description 25
- 230000005540 biological transmission Effects 0.000 claims description 23
- 238000000034 method Methods 0.000 claims description 22
- 230000037230 mobility Effects 0.000 claims description 19
- 239000007788 liquid Substances 0.000 claims description 17
- 238000010438 heat treatment Methods 0.000 claims description 14
- ARUVKPQLZAKDPS-UHFFFAOYSA-L copper(II) sulfate Chemical compound [Cu+2].[O-][S+2]([O-])([O-])[O-] ARUVKPQLZAKDPS-UHFFFAOYSA-L 0.000 claims description 12
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 claims description 11
- 238000011534 incubation Methods 0.000 claims description 11
- 229910000365 copper sulfate Inorganic materials 0.000 claims description 10
- 230000001276 controlling effect Effects 0.000 claims description 8
- 238000005406 washing Methods 0.000 claims description 8
- 238000004458 analytical method Methods 0.000 claims description 7
- 230000008859 change Effects 0.000 claims description 7
- 230000008569 process Effects 0.000 claims description 7
- 238000004364 calculation method Methods 0.000 claims description 3
- 239000012153 distilled water Substances 0.000 claims description 3
- 238000013507 mapping Methods 0.000 claims description 3
- 238000010606 normalization Methods 0.000 claims description 3
- 230000000149 penetrating effect Effects 0.000 claims description 3
- 230000000737 periodic effect Effects 0.000 claims description 3
- 230000001105 regulatory effect Effects 0.000 claims description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 3
- 238000009736 wetting Methods 0.000 claims description 3
- MQRJBSHKWOFOGF-UHFFFAOYSA-L disodium;carbonate;hydrate Chemical compound O.[Na+].[Na+].[O-]C([O-])=O MQRJBSHKWOFOGF-UHFFFAOYSA-L 0.000 claims 1
- 238000004659 sterilization and disinfection Methods 0.000 abstract description 16
- 238000002331 protein detection Methods 0.000 abstract description 4
- 230000035945 sensitivity Effects 0.000 description 19
- 238000010586 diagram Methods 0.000 description 8
- 230000006872 improvement Effects 0.000 description 8
- OHJMTUPIZMNBFR-UHFFFAOYSA-N biuret Chemical compound NC(=O)NC(N)=O OHJMTUPIZMNBFR-UHFFFAOYSA-N 0.000 description 7
- 210000003128 head Anatomy 0.000 description 7
- 230000007774 longterm Effects 0.000 description 6
- 230000000694 effects Effects 0.000 description 4
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- 239000003086 colorant Substances 0.000 description 3
- 238000002156 mixing Methods 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- FEMOMIGRRWSMCU-UHFFFAOYSA-N ninhydrin Chemical compound C1=CC=C2C(=O)C(O)(O)C(=O)C2=C1 FEMOMIGRRWSMCU-UHFFFAOYSA-N 0.000 description 3
- ZWLUXSQADUDCSB-UHFFFAOYSA-N phthalaldehyde Chemical compound O=CC1=CC=CC=C1C=O ZWLUXSQADUDCSB-UHFFFAOYSA-N 0.000 description 3
- 238000002965 ELISA Methods 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- 229910000366 copper(II) sulfate Inorganic materials 0.000 description 2
- 238000013461 design Methods 0.000 description 2
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- 238000012545 processing Methods 0.000 description 2
- 238000004611 spectroscopical analysis Methods 0.000 description 2
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- 244000025254 Cannabis sativa Species 0.000 description 1
- 229920000742 Cotton Polymers 0.000 description 1
- 102000008100 Human Serum Albumin Human genes 0.000 description 1
- 108091006905 Human Serum Albumin Proteins 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
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- 238000004847 absorption spectroscopy Methods 0.000 description 1
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- 238000002798 spectrophotometry method Methods 0.000 description 1
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- 238000012795 verification Methods 0.000 description 1
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/77—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
- G01N21/78—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator producing a change of colour
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Abstract
The invention discloses a detection device and a detection method for cleaning residual protein, and relates to the technical field of residual protein detection, wherein the detection device comprises a sampling assembly, and the sampling assembly comprises a sampling swab and a transparent reagent tube; a sealed diaphragm is arranged in the reagent tube, and a sealed reagent storage area is formed; the sample collecting end of the sampling swab can be inserted into the tube from the open end of the reagent tube and puncture the sealing diaphragm I to mix the collected sample with the detection reagent in the reagent storage area; the detection part comprises a constant temperature culture block, a photoelectric detection component and a controller; a constant temperature culture cavity is arranged in the constant temperature culture block, and test tube insertion holes and detection holes are formed in the top wall and the opposite side walls of the constant temperature culture block; the reagent tube can be inserted into the constant temperature culture cavity, and the outer wall of the reagent storage area corresponds to the detection hole; the transmitting end and the receiving end of the photoelectric detection assembly are respectively and correspondingly arranged at the positions of the outer side orifices of the two detection holes; the photoelectric detection component is electrically connected with the controller. The invention can be suitable for daily field detection operation of a hospital disinfection supply center.
Description
Technical Field
The invention relates to the technical field of residual protein detection, in particular to a detection device and a detection method for cleaning residual protein.
Background
In the field of medical cleaning and disinfection, reusable medical instruments need to be thoroughly cleaned first, and then subsequent disinfection and sterilization can achieve better effects. The cleaned medical instrument can enter a disinfection and sterilization link after being verified to be qualified. Whether the cleaning is thorough or not is always observed by naked eyes for a long time, and a residual protein detection method is invented by people later, and in the medical industry standard of YY/T0734.1-2018, ninhydrin, phthalic aldehyde and biuret are recommended to be used as three detection reagents of residual proteins.
In the ninhydrin method, as long as the color of the swab is artificially observed to become purple, it is confirmed that the residual protein is detected; in the phthalaldehyde method, spectrophotometry is required to detect fluorescent alkylthio-2-alkyl isoindole products at 340nm, and if the absorbance value is less than 0.02, the residual protein pollutants are less; when the biuret method is used, the color of the reagent becomes dark purple when the protein is present, the color is dark or light depending on the amount of the residual protein, and when the protein is not present, the reagent is apple green, and the approximate range of the residual protein can be judged by a color chart. The three standard methods described above each have disadvantages: the phthalaldehyde method requires the use of a spectrophotometer, and is not suitable for daily operation of a hospital disinfection supply center; the ninhydrin method and the biuret method rely on the color reaction of the protein to detect the protein residue, but the human eyes are required to judge the result, and the accuracy and the sensitivity have problems.
It is also feasible to detect residual proteins by using ultraviolet rays and other wavebands which cannot be perceived by the human eyes, for example Zhu Yuanrong, in the paper "ultraviolet absorption spectroscopy integration method for analyzing protein concentration" (spectroscopy and spectroscopic analysis, 2013, 7 th period), an absorption peak area integration method is proposed in the wavebands of 252nm-305nm, so as to solve the problem that the conventional ultraviolet absorption method cannot directly and accurately measure protein concentration value at 280 nm. Although this method can achieve a sensitivity of 20. Mu.g/ml, the use of a spectrophotometer is required by the ultraviolet method and is not suitable for the daily operation of a hospital disinfection supply center. The invention patent 'a detection method of human serum albumin residue with high sensitivity' (application publication number: CN 115524497A) is an ELISA detection method, which requires a special ELISA instrument, has a complex operation process, and is not suitable for daily operation of a hospital disinfection supply center.
Therefore, how to provide a detection device and a detection method for cleaning residual protein, which are suitable for daily on-site operation and have high sensitivity, aiming at the needs of medical instrument cleaning effect verification of a hospital disinfection supply center is a problem which needs to be solved by those skilled in the art.
Disclosure of Invention
In view of the above, the present invention provides a detection device and a detection method for cleaning residual proteins, which aim to solve the above technical problems. The detection device and the detection method for cleaning the residual protein provided by the invention can be suitable for daily on-site detection operation of a hospital disinfection supply center and have higher detection sensitivity.
In order to achieve the above purpose, the present invention adopts the following technical scheme:
in one aspect, the present invention provides a detection device for cleaning residual protein, comprising:
the sampling assembly comprises a sampling swab and a transparent reagent tube; a sealed diaphragm, namely a sealed reagent storage area, is arranged in the reagent tube; the sample collection end of the sampling swab can be inserted into the tube from the open end of the reagent tube and pierces the sealing diaphragm I so as to mix the collected sample with the detection reagent in the reagent storage area to form a reaction solution;
the detection part comprises a constant temperature culture block, a photoelectric detection component and a controller; a constant temperature culture cavity is arranged in the constant temperature culture block, and a test tube insertion hole and a penetrating detection hole are respectively formed in the top wall and the opposite side walls of the constant temperature culture block corresponding to the constant temperature culture cavity; the reagent tube can be inserted into the constant temperature culture cavity from the test tube insertion hole, and the outer wall of the reagent storage area corresponds to the detection hole; the transmitting end and the receiving end of the photoelectric detection assembly are respectively and correspondingly arranged at the outer side orifice positions of the two detection holes so as to detect the reaction liquid in the reagent storage area; the photoelectric detection component is electrically connected with the controller.
Compared with the prior art, the detection device for cleaning residual protein provided by the invention can be known through the technical scheme; the reagent storage area of reagent pipe is used for depositing detection reagent, and reagent storage area is sealed through sealed diaphragm one, and during the use, utilize the sampling swab that has gathered the washing residual protein to insert in the pipe and puncture sealed diaphragm one, can realize the mixing of the washing residual protein sample of taking and detection reagent, forms the reaction liquid in order to carry out the reaction of discolouring. During detection, the reagent tube is inserted into the constant temperature culture cavity of the constant temperature culture block, and the reagent storage area corresponds to the detection hole; the constant temperature culture cavity can culture the reaction liquid at constant temperature, so that the detection sensitivity is improved; the light emitted by the emitting end of the photoelectric detection component can penetrate through the transparent reagent tube and acts on the reaction liquid in the reagent storage area, the reaction liquid absorbs and filters part of the light waves to generate light to be detected, the light to be detected penetrates through the tube wall and is output to the receiving end of the photoelectric detection component, and the light to be detected is converted into an electric signal and then is transmitted to the controller for collection and processing. The detection reagent is stored in the transparent reagent tube in a sealing way, so that the use is convenient, and the reaction liquid does not need to be taken out during measurement; the reagent tube is directly put into the constant temperature culture block to finish detection, so that the detection device is convenient to operate and high in sensitivity, and can be suitable for daily field detection operation of a hospital disinfection supply center.
As a further improvement of the technical scheme, a second sealing diaphragm is arranged in the reagent tube, and the second sealing diaphragm is positioned at one side of the first sealing diaphragm away from the open end of the reagent tube at intervals so as to divide the reagent storage area into a closed reagent storage area A and a closed reagent storage area B; the sampling swab can be inserted into the tube from the open end of the reagent tube and sequentially pierce the first sealing diaphragm and the second sealing diaphragm downwards, and the sample at the sample collecting end is mixed with two detection reagents in the reagent storage area A and the reagent storage area B to form a reaction liquid.
The reagent storage A area and the reagent storage B area can be used for separately sealing and storing two different detection reagents, so that the detection reagents can be prevented from being combined in advance, the long-term storage of the detection reagents is facilitated, the complicated operation of configuring the mixed detection reagents is omitted, and the convenience of daily field detection operation is improved.
As a further improvement of the technical scheme, the sampling swab comprises a swab rod, a sampling head, a sliding sealing plug and a handle;
the sampling head is fixed at one end of the swab rod, and the free end of the sampling head is a sample collecting end; one end of the sliding sealing plug is fixed at the other end of the swab rod; the handle is fixed at the other end of the sliding sealing plug; the sampling head can be inserted into the tube from the open end of the reagent tube, and the sliding sealing plug can be in sliding sealing connection with the reagent tube.
The swab rod and the sampling head are hermetically stored in the reagent tube by the sliding sealing plug to form a sampling swab storage area, and the sampling swab storage area, the reagent storage area A and the reagent storage area B are mutually independent storage spaces, so that the drying and sterile state of the sampling head are realized, and the long-term storage of the sampling swab is facilitated; during sampling, the sampling swab can be pulled out through the handle, after the sampling is completed, the sampling swab can be inserted back into the reagent tube, and the sliding sealing plug can seal the open end of the reagent tube, so that the reagent tube is prevented from being polluted by external dirt and impurities, and the detection accuracy is improved; the sampling head can be driven to puncture the first sealing diaphragm and the second sealing diaphragm by pushing the handle, and the operation is convenient.
As a further improvement of the technical scheme, the test tube insertion hole is a guiding and positioning hole of the reagent tube; the bottom end of the reagent tube can penetrate through the test tube insertion hole so as to align the reagent storage area with the detection hole;
the bottom of the constant temperature culture block is provided with a constant temperature heating plate, and the side part of the constant temperature culture block is provided with a temperature sensor corresponding to the constant temperature culture cavity; the constant temperature heating plate and the temperature sensor are electrically connected with the controller.
During detection operation, the lower end of the reagent tube can be inserted into the jack, the jack plays a role in guiding and positioning the reagent tube, and the bottom end of the reagent tube can be abutted against the bottom wall of the constant-temperature culture cavity, so that the reagent storage area is accurately aligned to the detection hole, the positioning precision of the reagent tube is improved, and the reliability and the accuracy of detection are further improved; the controller controls the constant temperature heating plate to heat, and the temperature sensor feeds back the temperature of the constant temperature culture cavity, so that the constant temperature regulation of the constant temperature culture cavity can be realized, the constant temperature regulation of the temperature is provided for the color reaction, and the sensitivity and the precision of detection are improved.
As a further improvement of the above technical solution, the photodetection assembly includes a light emitting plate and a receiving plate;
the light-emitting plate and the receiving plate are symmetrically fixed on two side walls of the constant temperature culture block; a light source is fixed on the side wall of the light-emitting plate, which is close to the constant-temperature culture block, a photoelectric sensor is arranged on the side wall of the receiving plate, which is close to the constant-temperature culture block, and the light source and the photoelectric sensor are respectively positioned at two ends in the detection hole; the light source and the photoelectric sensor are electrically connected with the controller.
The light beam emitted by the light source can penetrate through the constant temperature culture cavity through the detection hole, and when the reagent tube to be detected is placed in the constant temperature culture cavity, the light beam emitted by the light source can penetrate through the outer wall of the transparent reagent tube and acts on the reaction liquid; the photoelectric sensor is used for receiving light to be detected transmitted out of the reaction liquid and performing photoelectric conversion on the light to be detected.
As a further improvement of the above technical solution, the light source is a white light source; the photoelectric sensor comprises a plurality of photoelectric receiving tubes, and the plurality of photoelectric receiving tubes are arranged in parallel to form an array type receiving end; the front end of each photoelectric receiving tube is provided with an optical filter; the optical filters respectively correspond to different preset light transmission wave bands; the photoelectric receiving tubes are electrically connected with the controller to receive and feed back the filtered light signals of the appointed wave bands.
The light source is a white light source and comprises visible light waves of various wave bands; the receiving end formed by a plurality of photoelectric receiving tubes distributed in an array manner can receive and convert the light wave signals; the front end of each photoelectric receiving tube is covered with an independent optical filter, and each optical filter can correspond to different visible light transmission wave bands, namely, the full-wave white light can be divided into a plurality of wave band intervals with different colors for sub-wave band detection; the photoelectric receiving tubes can respectively detect the light to be detected of different preset wave bands; the balance coverage of the color interval of the reaction liquid can be realized, and the detection precision is improved.
As a further improvement of the technical scheme, the reagent storage area A stores anhydrous sodium carbonate, and the reagent storage area B stores copper sulfate; or copper sulfate is stored in the reagent storage area A, and anhydrous sodium carbonate is stored in the reagent storage area B.
The invention uses biuret reaction as a basis to construct a detection reagent, and anhydrous sodium carbonate and copper sulfate are placed in an independent and closed reagent storage area, so that the technical problem that the reaction solution is combined in advance and is inconvenient for long-term storage is solved; the integrated design of the device structure is realized, and the technical effects of simple structure, suitability for daily rapid operation and meeting the use requirement of a disinfection supply center are achieved.
The invention also provides a detection method for cleaning residual protein, which comprises the detection device for cleaning residual protein, and comprises the following steps:
step one: wiping the surface of the cleaned detected instrument by using the head of the sampling swab so as to collect residual protein on the detected instrument;
step two: inserting the sampling swab head into the tube from the top end of the reagent tube and downwards puncturing the sealing diaphragm I, and enabling the sampling swab head to enter the reagent storage area so as to fully mix and contact the sampled residual protein sample with the detection reagent in the reagent storage area to form a reaction liquid;
step three: pulling up the sampling swab and enabling the head of the sampling swab to leave the reagent storage area, and inserting the part of the reagent tube corresponding to the reagent storage area into a constant temperature culture cavity for constant temperature culture of reaction liquid so as to perform color change reaction;
Step four: the controller is used for controlling the emitting end of the photoelectric detection component to emit optical signals, the optical signals pass through the detection hole and penetrate through the reagent storage area to be received by the receiving end of the photoelectric detection component, and the photoelectric detection component is used for converting the received optical signals into electric signals to be transmitted and fed back to the controller;
step five: and the controller processes the electric signals fed back by the photoelectric detection assembly by adopting a weighted mobility analysis method, and finally, the quantitative value of the cleaning residual protein is obtained.
As a further improvement of the above technical solution, the specific steps for collecting the residual protein on the detected instrument in the first step are as follows: holding the handle, pulling out the sampling swab from the reagent tube, wetting the sampling head of the sampling swab with distilled water, manually operating the sampling head to wipe the surface of the detected instrument, and collecting cleaning residues on the surface of the detected instrument;
the specific steps of the second step are as follows: reinserting the sampled sampling swab back into the reagent tube, firstly inserting the sampling head from the opening at the top end of the reagent tube, pushing the handle, and sequentially puncturing the sealing diaphragm I and the sealing diaphragm II by using the sampling head to enable the sampling head to enter the reagent storage area A, and finally fully mixing and contacting anhydrous sodium carbonate and copper sulfate reagent in the reagent storage area A, and shaking the test tube forcefully to enable collected cleaning residues to fall off from the sampling head and remain the reagent storage area A for participating in color change reaction;
The specific steps of the third step are as follows: preheating the constant-temperature culture cavity, controlling and starting the constant-temperature heating plate through the controller, and regulating and controlling the temperature of the constant-temperature culture cavity to be 60 ℃ and keeping unchanged; and inserting the reagent tube with the collected cleaning residual protein into the constant temperature culture cavity through the insertion hole, so that the bottom end of the reagent tube is abutted with the bottom end of the constant temperature culture cavity, and the reagent storage area A is ensured to be aligned with the detection hole for constant temperature culture.
The specific steps of the fourth step are as follows: the controller is used for turning on the light source, the emitted white light irradiates and penetrates through the corresponding test tube wall of the reagent storage area A, and the white light is output through the test tube wall after being absorbed and filtered by the reaction liquid in the reagent storage area A, so that the white light reaches the photoelectric sensor; the controller performs periodic data acquisition on a plurality of photoelectric receiving tubes; the data collected in each period is D (i, t), where i=1, 2,3 … N, t=1, 2,3 … N, where i is the number of the photoelectric receiving tubes, N is the number of the photoelectric receiving tubes, the light transmission bands different from the plurality of optical filters are in one-to-one correspondence, the band numbers of the light transmission bands are Ch1, ch2, ch3 … Chn, t are sampling moments, the sampling moments are numbered sequentially from 1, and N is the moment number corresponding to the last sampling value.
The specific steps of the fifth step are as follows: firstly, carrying out weighted normalization on the sample value of each photoelectric receiving tube; determining each weighted value omega according to the gain of the photoelectric receiving tube and the attenuation value of the optical filter i I=1, 2,3 … n; each sample value is multiplied by a corresponding weighting value to obtain a normalized sample value:
σ i （t）=ω i x D (i, t), wherein i=1, 2,3 … n; t=1, 2,3 … N;
secondly, normalizing the Bai Guangguang source intensity; in the case of initially inserting the reagent tube and turning on the white light source, the data of each photoelectric receiving tube is sampled once, and the weighted sampling value in the case of initially inserting the reagent tube is represented by t=0, the normalized coefficient λ of the white light source intensity is represented as:
；
then, calculating the band mobility of each photoelectric receiving tube; band mobility S i The calculation formula of (2) is as follows:
，i=1，2，3…n-1；
d i is the distance between the central wavelength of the light transmission wave band corresponding to the photoelectric receiving tube with the number i and the central wavelength of the light transmission wave band with the wave band number Chn; band mobility is the full path delta of each transmission band relative to the transmission band with band number Chn since the start of incubation;
finally, the band mobility of all the photoelectric receiving tubes is accumulated, and the normalized coefficient of the white light source intensity is considered, so that the residual protein reading R (n) at the current moment can be obtained, which is expressed as:
。
As a further improvement of the technical scheme, an alarm threshold is set for R (n) according to the requirement; at any time of incubation, when the value of R (n) exceeds the alarm threshold, indicating that the residual protein reading has exceeded the expected value, the early end of the incubation process can be controlled; when the cultivation is finished, the controller displays the reading value of the detection device;
by preparing the tested object with known protein quantity, a detection curve of the mapping relation between the reading of the detection device and the residual protein quantity can be obtained. The reading value of the detection device can be converted into the amount of the washing residual protein in μg/ml according to the detection curve every time a sample of unknown residual protein amount is tested.
Compared with the prior art, the invention discloses a detection device for cleaning residual protein, which has the following advantages and beneficial effects:
1. the reagent tube is divided into a reagent storage area A, a reagent storage area B and a sampling swab storage area by the penetrable sealing diaphragm I and the sealing diaphragm II; the independent storage of two detection reagents and the sealed storage of a sampling swab are realized; when the device is used, the sampling swab is used for puncturing the first sealing diaphragm and the second sealing diaphragm to mix the collected sample with the detection reagent, so that the device has the technical effects of simple structure, convenience in operation and applicability to daily field detection operation of a hospital disinfection supply center.
2. The detection device does not need to take out the reaction liquid during the measurement operation; the reagent tube can be directly placed in the constant temperature culture block to complete constant temperature culture and detection, and the detection operation is convenient and fast, and the reliability and the sensitivity are high.
3. The detection device provided by the invention can realize balanced coverage of the color interval of the reaction liquid and improve the detection precision by dividing the full-band white light into a plurality of band intervals with different colors for sub-band photoelectric detection.
4. On the basis of the detection device, the data acquired by the photoelectric detection assembly is analyzed and processed by adopting a weighted mobility analysis method, so that the quantitative value of the cleaning residual protein is finally obtained, and the detection sensitivity is greatly improved.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings that are required to be used in the embodiments or the description of the prior art will be briefly described below, and it is obvious that the drawings in the following description are only embodiments of the present invention, and that other drawings can be obtained according to the provided drawings without inventive effort for a person skilled in the art.
FIG. 1 is a schematic diagram of a sampling swab structure of a detection device for cleaning residual proteins;
FIG. 2 is a schematic diagram of a sample swab structure with a scraping assembly of the detection device for cleaning residual proteins according to the present invention;
FIG. 3 is a schematic diagram showing the overall structure of a detection device for cleaning residual proteins according to the present invention;
FIG. 4 is a schematic view showing another view of the whole structure of a device for detecting protein residues in cleaning according to the present invention;
FIG. 5 is a schematic diagram showing the assembly of a photoelectric detection assembly and a constant temperature culture block of a detection device for cleaning residual proteins;
FIG. 6 is a schematic diagram of the structure of a photoelectric sensor of a detection device for cleaning residual protein;
FIG. 7 is a schematic diagram showing the structure of a liquid scraping assembly of a detection device for cleaning residual proteins;
FIG. 8 is a schematic diagram showing a detection sensitivity curve of a detection method for cleaning residual proteins according to the present invention;
in the figure: 1. sampling a swab; 11. a swab stem; 12. a sampling head; 13. a handle; 14. sliding the sealing plug; 2. a reagent tube; 21. sealing the first diaphragm; 22. a reagent storage area; 221. a reagent storage area A; 222. a reagent storage area B; 23. sealing the second diaphragm; 24. a sampling swab storage area; 3. a constant temperature culture block; 31. a constant temperature culture cavity; 32. a detection hole; 33. a jack; 34. a constant temperature heating plate; 35. a temperature sensor; 4. a photodetection assembly; 41. a light emitting panel; 411. a light source; 42. a receiving plate; 421. a photoelectric sensor; 4211. a photoelectric receiving tube; 4212. a light filter; 5. a controller; 6. a liquid scraping assembly; 61. a liquid scraping elastic sheet.
Detailed Description
Embodiments of the present invention are described in detail below, examples of which are illustrated in the accompanying drawings, wherein like or similar reference numerals refer to like or similar elements or elements having like or similar functions throughout. The embodiments described below by referring to the drawings are illustrative and intended to explain the present invention and should not be construed as limiting the invention.
In the description of the present invention, it should be understood that the terms "upper", "lower", "front", "rear", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer", etc. indicate orientations or positional relationships based on the drawings, are merely for convenience in describing the present invention and simplifying the description, and do not indicate or imply that the devices or elements referred to must have a specific orientation, be configured and operated in a specific orientation, and thus should not be construed as limiting the present invention.
Furthermore, the terms "first," "second," and the like, are used for descriptive purposes only and are not to be construed as indicating or implying a relative importance or implicitly indicating the number of technical features indicated. Thus, a feature defining "a first" or "a second" may explicitly or implicitly include one or more such feature. In the description of the present invention, the meaning of "a plurality" is two or more, unless explicitly defined otherwise.
In the present invention, unless explicitly specified and limited otherwise, the terms "mounted," "connected," "secured," and the like are to be construed broadly, and may be, for example, fixedly connected, detachably connected, or integrally formed; can be mechanically or electrically connected; can be directly connected or indirectly connected through an intermediate medium, and can be communicated with the inside of two elements or the interaction relationship of the two elements. The specific meaning of the above terms in the present invention can be understood by those of ordinary skill in the art according to the specific circumstances.
As shown in fig. 1 to 7, the present embodiment provides a detection apparatus for cleaning residual protein, including:
the sampling assembly comprises a sampling swab 1 and a transparent reagent tube 2; a first sealing diaphragm 21 is arranged in the reagent tube 2, so that a sealed reagent storage area 22 is defined below the first sealing diaphragm 21 in the reagent tube 2; the sample collection end of the sampling swab 1 can be inserted into the tube from the open end of the reagent tube 2 and the sealing diaphragm 21 is pierced to mix the collected sample with the detection reagent in the reagent storage area 22 to form a reaction solution;
the detection part comprises a constant temperature culture block 3, a photoelectric detection component 4 and a controller 5; the inside of the constant temperature culture block 3 is provided with a constant temperature culture cavity 31, and the top wall and the opposite side walls of the constant temperature culture block 3 are respectively provided with a test tube insertion hole 33 and a penetrating detection hole 32 corresponding to the constant temperature culture cavity 31; the reagent tube 2 can be inserted into the constant temperature culture chamber 31 from the test tube insertion hole 33, and the outer wall of the reagent storage area 22 corresponds to the detection hole 32; the transmitting end and the receiving end of the photoelectric detection assembly 4 are respectively correspondingly arranged at the outer orifice positions of the two detection holes 32 so as to detect the reaction liquid in the reagent storage area 22; the photodetection module 4 is electrically connected with the controller 5.
The embodiment provides a detection device for cleaning residual protein; the reagent storage area 22 of the reagent tube 2 is used for storing detection reagents, the reagent storage area 22 is closed by the first sealing diaphragm 21, and when the reagent storage area 22 is used, the collected cleaning residual protein sample swab 1 is inserted into the tube and the first sealing diaphragm 21 is pierced, so that the collected cleaning residual protein sample and the detection reagents can be mixed to form a reaction liquid for color change reaction. During detection, the reagent tube 2 is inserted into the constant temperature culture cavity 31 of the constant temperature culture block 3, and the reagent storage area 22 corresponds to the detection hole 32; the constant temperature culture cavity 31 can perform constant temperature culture on the reaction liquid, so that the detection sensitivity is improved; the light emitted from the emitting end of the photoelectric detection component 4 can penetrate through the transparent reagent tube 2 and acts on the reaction liquid in the reagent storage area 22, the reaction liquid absorbs and filters part of the light waves to generate light to be detected, the light to be detected penetrates through the tube wall and is output to the receiving end of the photoelectric detection component 4, and the light to be detected is converted into an electric signal and then transmitted to the controller 5 for collection and processing. The detection reagent is stored in the transparent reagent tube 2 in a sealing way, so that the use is more convenient, and the reaction liquid does not need to be taken out during the measurement; the reagent tube 2 is directly put into the constant temperature culture block 3 to finish detection, so that the operation is convenient and quick, the sensitivity is high, and the method is suitable for daily field detection operation of a hospital disinfection supply center.
In some embodiments, the inside of the constant temperature culture block 3 is a hollow structure, and the hollow part forms a culture chamber of the reaction liquid; the number of the thermostatic culture chambers 31 inside the thermostatic culture block 3 can be plural; the plurality of constant temperature culture chambers 31 are arranged at intervals along the length direction of the constant temperature culture block 3; a detection hole 32 is arranged corresponding to each constant temperature culture block 3; the photodetection assembly 4 is provided with a plurality of emitting ends and receiving ends and is provided corresponding to the plurality of detection holes 32, respectively. The plurality of constant temperature culture chambers 31 can meet the requirement of simultaneous detection of a plurality of reagent tubes 2, and the detection efficiency is increased.
In some embodiments, a second sealing membrane 23 is arranged in the reagent tube 2, and the second sealing membrane 23 is spaced on one side of the first sealing membrane 21 away from the open end of the reagent tube 2 so as to divide the reagent storage area 22 into a closed reagent storage area A221 and a reagent storage area B222; the sampling swab 1 can be inserted into the tube from the open end of the reagent tube 2 and sequentially pierce the first sealing diaphragm 21 and the second sealing diaphragm 23 downwards, and the sample at the sample collection end is mixed with two detection reagents in the reagent storage A area 221 and the reagent storage B area 222 to form a reaction solution.
The reagent storage A area 221 and the reagent storage B area 222 can separate two different detection reagents for sealing storage, can prevent the detection reagents from being combined in advance, are convenient for long-term storage of the detection reagents, save complicated operation of configuring mixed detection reagents, and improve convenience of daily field detection operation.
In some embodiments, the sampling swab 1 comprises a swab stem 11, a sampling head 12, a sliding sealing plug 14, and a handle 13;
the sampling head 12 is fixed at one end of the swab rod 11, and the free end of the sampling head is a sample collecting end; one end of the sliding sealing plug 14 is fixed at the other end of the swab rod 11; a handle 13 is fixed at the other end of the sliding seal plug 14; the sampling head 12 can be inserted into the reagent vessel 2 from its open end, and the sliding sealing plug 14 can be in sliding sealing connection with the reagent vessel 2. A closed sampling swab storage area 24 is formed between the sealing diaphragm one 21 and the sliding sealing plug 14 in the reagent tube 2; the handle 13 is positioned outside the reagent vessel 2; pushing handle 13 may drive sampling head 12 to pierce sealing diaphragm one 21 and sealing diaphragm two 23 in sequence.
Specifically, the first sealing diaphragm 21 and the second sealing diaphragm 23 are plastic films, the inner wall of the reagent tube 2 is provided with steps, and the first sealing diaphragm 21 and the second sealing diaphragm 23 are adhered and sealed at the top ends of the corresponding steps in the reagent tube 2.
The swab rod 11 and the sampling head 12 are hermetically stored in a sampling swab storage area 24 of the reagent tube 2 by the sliding sealing plug 14, and the sampling swab storage area 24, the reagent storage A area 221 and the reagent storage B area 222 are mutually independent storage spaces, so that the drying and sterile state of the sampling head 12 is realized, and the long-term storage of the sampling swab is facilitated; during sampling, the sampling swab 1 can be pulled out through the handle 13, and can be inserted back into the reagent tube 2 after the sampling is completed, and the sliding sealing plug 14 can seal the open end of the reagent tube 2, so that the reagent tube 2 is ensured not to be polluted by external dirt and impurities, and the detection accuracy is improved; the sampling head 12 can be driven to puncture the sealing diaphragm I21 and the sealing diaphragm II 23 by pushing the handle 13, so that the operation is convenient.
In particular, the reagent vessel 2 may be made of transparent plastic or glass; the reagent vessel 2 is required to have very low absorption of visible light, allowing visible light to penetrate into the vessel wall and out. The swab rod 11 is made of a mixture of ABS and PC plastics, so that the swab rod 11 has hardness and flexibility. The handle 13 has a multi-layer concave-convex structure and an anti-skid function, so that the swab rod 11 is conveniently pulled out during use; the handle 13 and the sliding sealing plug 14 can be of an integrated structure and made of rubber materials; the outer diameter of the handle 13 is smaller than the outer diameter of the sliding sealing plug 14; the outside diameter of the sliding seal plug 14 is slightly larger than the inside diameter of the reagent vessel 2, so that the sliding seal plug 14 is slidably sealed in the reagent vessel 2 to maintain a clean state inside the reagent vessel 2.
The head of the swab rod 11 with flexibility is wrapped by cotton to form a sampling head 12, and the wrapping material for manufacturing the sampling head has enough flexibility, so that the sampling swab 1 can adapt to the complex surface structure of a sampled instrument, and stretches into gaps and corners difficult to sample to finish the sampling of residual proteins; when the sample is finished and the color reaction is needed, the swab rod 11 with certain hardness is pushed downwards through the handle 13 and then goes deep into the reagent tube 2 for a certain distance, so that the sealing membrane can be punctured, and the sampling head 12 and two different detection reagents can be fully contacted and mixed.
In some embodiments, a wiper assembly 6 is also included; the liquid scraping assembly 6 consists of a plurality of liquid scraping elastic sheets 61; the plurality of scraping spring plates 61 are uniformly arranged along the circumferential direction of the inner wall of the reagent tube 2, are positioned on the side of the first sealing diaphragm 21, which is far away from the second sealing diaphragm 23, and are positioned below the sampling head 12; one end of each of the plurality of liquid scraping elastic pieces 61 is adhered and fixed with the inner wall of the reagent tube 2, the other end of each of the plurality of liquid scraping elastic pieces 61 is a free end, and the plurality of liquid scraping elastic pieces 61 are obliquely arranged downwards; the diameter of a central hole formed by the free end of the plurality of scraping elastic pieces 61 is smaller than the outer diameter of the sampling head 12; the scraping spring piece 61 is a rubber sheet or a plastic sheet with certain elasticity; when the sampling head 12 moves away from the reagent storage area B222, the sampling head 12 moves downwards smoothly through the scraping elastic pieces 61, and the plurality of scraping elastic pieces 61 bend reversely to jointly squeeze the sampling head 12, so that the reaction liquid adsorbed on the surface of the sampling head can be extruded out and flows back to the reagent storage area B222 and the reagent storage area A221; when the sampling head 12 moves upwards to separate from the plurality of liquid scraping spring pieces 61, the liquid scraping spring pieces 61 can rebound the extruded reaction liquid downwards to scatter the reaction liquid, so that the adsorption quantity of the sampling head 12 to the reaction liquid can be reduced, and the detection precision is improved.
In some embodiments, the test tube insertion hole 33 is a guide positioning hole of the reagent tube 2; the bottom end of the reagent tube 2 may penetrate through the test tube insertion hole 33 so that the reagent storage area 22 is aligned with the detection hole 32;
The bottom of the constant temperature culture block 3 is fixed with a constant temperature heating plate 34, and the side part of the constant temperature culture block 3 is provided with a temperature sensor 35 corresponding to the constant temperature culture cavity 31; the constant temperature heating plate 34 and the temperature sensor 35 are electrically connected with the controller 5.
During detection operation, the lower end of the reagent tube 2 can be inserted into the insertion hole 33, the insertion hole 33 plays a role in guiding and positioning the reagent tube 2, and the bottom end of the reagent tube 2 can be abutted against the bottom wall of the constant temperature culture cavity 31, so that the reagent storage area 22 is accurately aligned with the detection hole 32, the positioning precision of the reagent tube 2 is improved, and the reliability and accuracy of detection are further improved; the constant temperature heating plate 34 is controlled by the controller 5 to heat, the temperature of the constant temperature culture cavity 31 is fed back by the temperature sensor 35, the constant temperature regulation and control of the constant temperature culture cavity 31 can be realized, the constant temperature regulation of the temperature is provided for the color reaction, and the sensitivity and the precision of detection are improved.
In some embodiments, the photodetection assembly 4 comprises a light emitting plate 41 and a receiving plate 42;
the light-emitting plate 41 and the receiving plate 42 are symmetrically fixed on two side walls of the constant temperature culture block 3 and close two end orifices of the detection hole 32; a light source 411 is fixed on the side wall of the light-emitting plate 41, which is close to the constant temperature culture block 3, a photoelectric sensor 421 is fixed on the side wall of the receiving plate 42, which is close to the constant temperature culture block 3, and the light source 411 and the photoelectric sensor 421 are respectively positioned at two ends in the detection hole 32; the light source 411 and the photoelectric sensor 421 are electrically connected to the controller 5. The temperature sensor 35 is fixed to the side wall of the light-emitting plate 41 near the thermostatic incubator block 3.
The light beam emitted by the light source 411 can penetrate through the constant temperature culture cavity 31 through the detection hole 32, and when the reagent tube 2 to be detected is placed in the constant temperature culture cavity 31, the light beam emitted by the light source 411 can penetrate through the outer wall of the transparent reagent tube 2 and act on the reaction liquid; the photoelectric sensor 421 is configured to receive light to be detected transmitted from the reaction solution, and perform photoelectric conversion on the light to be detected; the light emitting plate 41 and the receiving plate 42 close the two end apertures of the detection hole 32 to prevent external light from interfering with photoelectric measurement.
In some embodiments, light source 411 is a white light source; the photoelectric sensor 421 includes a plurality of photoelectric receiving tubes 4211, and the plurality of photoelectric receiving tubes 4211 are parallel and arranged at intervals to form an array-type receiving end; the front end of each photoelectric receiving tube 4211 is provided with an optical filter 4212; the plurality of optical filters 4212 respectively correspond to different preset light transmission wave bands; the plurality of photoelectric receiving tubes 4211 are electrically connected to the controller 5 to receive and feed back the filtered optical signals of the specified wavelength band.
The light source 411 is a 6500K white light source, which contains visible light waves of various wave bands; the receiving end formed by the plurality of photoelectric receiving tubes 4211 distributed in an array manner can receive and convert the light wave signals; the front end of each photoelectric receiving tube 4211 is covered with an independent optical filter 4212, and each optical filter 4212 can correspond to different visible light transmission wave bands, namely, the full-wave band white light can be divided into a plurality of wave band intervals with different colors for sub-wave band detection; the photoelectric receiving tubes 4211 can respectively detect the light to be detected of different preset wave bands; the balance coverage of the color interval of the reaction liquid can be realized, and the detection precision is improved.
Specifically, the photoelectric receiving tube 4211 of the present embodiment is selected from photoelectric receiving tubes; six photodiodes are adopted at the receiving end of the photoelectric sensor 421, the corresponding band numbers of the photodiodes are Ch1, ch2, ch3, ch4, ch5 and Ch6, the corresponding wavelength ranges of each band are 405-425nm, 435-455nm, 470-490nm, 505-525nm, 545-565nm and 580-600nm, and the typical area from blue to yellow is covered. The six wavelength bands are selected, the interval width of the wavelength is 20nm, the interval between the wavelength bands is 15nm, the detection sensitivity of the photodiode and the attenuation passband width of the optical filter are both considered, and the balanced coverage of the color interval of the reaction liquid is realized.
In some embodiments, reagent storage a zone 221 stores anhydrous sodium carbonate and reagent storage B zone 222 stores copper sulfate; or the reagent storage a area 221 stores copper sulfate, and the reagent storage B area 222 stores anhydrous sodium carbonate.
Specifically, the temperature of the thermostatic incubation cavity 31 may be set to a thermostatic value of 60 degrees celsius; can also be set by the user according to the requirement. The program on the controller 5 can complete the operations of emitting white light source, receiving data of the photodiode, heating control of the constant temperature heating plate, acquiring data of the temperature sensor, and the like. The controller 5 can realize the determination of the biuret reaction result under the action of the detection algorithm by acquiring the data of the photodiode.
The invention uses biuret reaction as base to construct detection reagent, which is typical anhydrous sodium carbonate and cupric sulfate solution in biuret reaction; the anhydrous sodium carbonate and the cupric sulfate are placed in an independent and closed reagent storage area, so that the technical problem that the reaction solution is combined in advance and is inconvenient for long-term storage is solved; the integrated design of the device structure is realized, the device is convenient to operate and low in price, the structure is simple, the device is suitable for daily rapid operation, the application requirement of a disinfection supply center can be met, and the device has higher residual protein detection sensitivity.
The invention also provides a detection method for cleaning residual protein, which comprises the following steps:
step one: wiping the surface of the cleaned detected instrument by the head of the sampling swab 1 to collect residual protein on the detected instrument;
step two: inserting a sample collection end of a sampling swab 1 for collecting a sample into the tube from the open end of the reagent tube 2, downwards puncturing the sealing diaphragm I21, and enabling the head of the sampling swab 1 to enter the reagent storage area 22 so as to fully mix and contact the collected residual protein sample with the detection reagent in the reagent storage area 22 to form a reaction solution;
Step three: the sampling swab 1 is pulled up, the sample collection end of the sampling swab leaves the reagent storage area 22, and the pipe wall section of the reagent pipe 2 corresponding to the reagent storage area 22 is inserted into the constant temperature culture cavity 31 for constant temperature culture of the reaction liquid so as to carry out color change reaction;
step four: the controller 5 controls the emitting end of the photoelectric detection component 4 to emit an optical signal, the optical signal passes through the detection hole 32 and penetrates through the reagent storage area 22 to be received by the receiving end of the photoelectric detection component 4, and the photoelectric detection component 4 converts the received optical signal into an electrical signal to be transmitted and fed back to the controller 5;
step five: the controller 5 processes the electric signal fed back by the photoelectric detection assembly 4 by adopting a weighted mobility analysis method, and finally obtains the quantitative value of the cleaning residual protein.
In some embodiments, the specific steps of collecting the residual protein on the tested device in the first step are: holding the handle 13, pulling out the sampling swab 1 from the reagent tube 2, wetting the sampling head 12 of the sampling swab 1 with distilled water, manually operating to wipe the sampling head 12 on the surface of the detected instrument, and collecting cleaning residues on the surface of the detected instrument;
the specific steps of the second step are as follows: reinserting the sampled sampling swab 1 into the reagent tube 2, firstly inserting the sampling head 12 from the top end opening of the reagent tube 2, pushing the handle 13 downwards, sequentially puncturing the sealing diaphragm I21 and the sealing diaphragm II 23 by using the sampling head 12, enabling the sampling head 12 to enter the reagent storage A area 221, enabling anhydrous sodium carbonate and copper sulfate reagent to be fully mixed and contacted in the reagent storage A area 221, and shaking the test tube forcefully to enable collected cleaning residues to fall off from the sampling head 12 and remain in the reagent storage A area 221 to participate in color change reaction; after full mixing, the sampling swab 1 needs to be pulled up, the sampling head 12 is separated from the reagent storage area A221, the handle 13 is pulled up and down, the reaction liquid adsorbed by the sampling head 12 is extruded and sprung by the plurality of liquid scraping spring pieces 61 of the liquid scraping assembly 6, and only the fully mixed reaction liquid and cleaning residues exist in the reagent storage area A221, so that shielding of the sampling head 12 to detection light is avoided.
The specific steps of the third step are as follows: preheating a constant-temperature culture cavity 31, controlling and starting a constant-temperature heating plate 34 through a controller 5, and regulating and controlling the temperature of the constant-temperature culture cavity 31 to be 60 ℃ and keeping unchanged; the reagent tube 2 for completing the collection of the cleaning residual protein is inserted into the constant temperature culture cavity 31 through the insertion hole 33, so that the bottom end of the reagent tube 2 is abutted with the bottom end of the constant temperature culture cavity 31, the reagent storage A area 221 is aligned to the detection hole 32, and the protein chromogenic reaction is carried out at the environmental temperature of 60 ℃.
The specific steps of the fourth step are as follows: the controller 5 turns on the light source 411, the emitted white light irradiates and penetrates the corresponding test tube wall of the reagent storage A area 221, the white light is absorbed and filtered by the reaction liquid in the reagent storage A area 221, and the generated light penetrates the test tube wall to be output and reaches the photoelectric sensor 421; the controller 5 performs periodic data acquisition on the plurality of photoelectric receiving tubes 4211; the sampling period is typically greater than 30 seconds; the data collected in each period is D (i, t), where i=1, 2,3 … N, t=1, 2,3 … N, where i is the number of the photoelectric receiving tubes 4211, N is the number of the photoelectric receiving tubes 4211, the multiple photoelectric receiving tubes 4211 are in one-to-one correspondence with different light transmission bands of the multiple optical filters 4212, and the band numbers of the light transmission bands are Ch1, ch2, ch3 … Chn; t is the time of sampling, the serial numbers are started from 1, and N is the time number corresponding to the last sampling value.
The specific steps of the fifth step are as follows: first, the sample value of each photoelectric receiving tube 4211 is weighted and normalized; each weighted value ω is determined based on the gain of the photo receiver tube 4211 and the attenuation value of the filter 4212 i I=1, 2,3 … n; each sample value is multiplied by a corresponding weighting value to obtain a normalized sample value:
σ i （t）=ω i x D (i, t), where i=1, 2,3 … N, t=1, 2,3 … N;
secondly, in order to eliminate the influence of the intensity of the white light source, the final algorithm output value has consistency in different detection devices, and the Bai Guangguang source intensity is normalized; in the case of initially inserting the reagent tube 2 and turning on the white light source, the data of each of the photo-receiving tubes 4211 is sampled once, and the weighted sampling value in the case of initially inserting the reagent tube 2 is represented by t=0, the normalized coefficient λ of the white light source intensity is represented as:
；
then, the band mobility of each photoelectric receiving tube 4211 is calculated; the color of the reaction liquid is grass green when the reaction liquid is initially mixed, if residual protein exists in the reaction liquid, the color of the reaction liquid gradually changes to blue, and the reaction liquid changes to purple when the amount of protein is large. Band mobility S i The calculation formula of (2) is as follows:
，i=1，2，3…n-1；
d i is the distance between the center wavelength of the light transmission band corresponding to the photoelectric receiving tube 4211 with the number i and the center wavelength of the light transmission band with the band number Chn; band mobility is the full path delta of each transmission band relative to the transmission band with band number Chn since the start of incubation;
Specifically, in this embodiment, the number of the photo-receiving tubes 4211 and the optical filters 4212 is six, and the wavelength ranges corresponding to each band are 405-425nm for Ch1, 435-455nm for Ch2, 470-490nm for Ch3, 505-525nm for Ch4, 545-565nm for Ch5, and 580-600nm for Ch 6. Six weighting values are ω, respectively, determined based on the gain of the photo receiver tube 4211 and the attenuation of the filter 4212 provided by the manufacturer 1 =2.12、ω 2 =1.98、ω 3 =1.57、ω 4 =1.39、ω 5 =1.12、ω 6 =1.00; the normalized sample enables sample values of different wave bands to be comparable; the shorter the wavelength band, the larger the weighting value that needs to be compensated. d, d i The distance between the center wavelength of the band with the number i and the center wavelength of the yellow band Ch6 is given, and N is the time number corresponding to the last sampling value. From the band division parameters Ch1, ch2, ch3, ch4, and Ch5, d is known 1 =175，d 2 =145，d 3 =110，d 4 =75，d 5 =35。
Finally, the band mobilities of all the photoreceptors 4211 are summed up and the current time residual protein reading R (n) can be obtained taking into account the normalized coefficient of white light source intensity, expressed as:
。
in some embodiments, an alarm threshold is set for R (n) as needed; at any time of incubation, when the value of R (n) exceeds the alarm threshold, indicating that the residual protein reading has exceeded the expected value, the early end of the incubation process can be controlled; when the incubation is finished, the controller 5 displays the reading value of the detection device;
In the weighted mobility analysis method, the intensity normalization of the white light source and the weighted value of each photodiode are taken into consideration, so that the detection device reading value is in direct proportion to the amount of the collected residual protein. By preparing the tested object with known protein quantity, a detection curve of the mapping relation between the reading of the detection device and the residual protein quantity can be obtained. Each time when a sample with unknown residual protein is tested, the reading value of a detection device can be converted into the cleaning residual protein in mug/ml according to a detection curve; the residual protein reading is a dimensionless number, the magnitude of which is linear with the residual protein amount, as shown in FIG. 8, and a schematic diagram of the detection sensitivity curve is shown in FIG. 8.
The minimum amount of residual protein detected that can be achieved by a proportional detection curve is the sensitivity of the weighted mobility assay. Through testing, the detection sensitivity of 3 mug/ml can be achieved by combining a weighted mobility analysis method based on the continuous analysis of the color sampling data of the reaction liquid by the detection device.
In the description of the present specification, a description referring to terms "one embodiment," "some embodiments," "examples," "specific examples," or "some examples," etc., means that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the present invention. In this specification, schematic representations of the above terms are not necessarily directed to the same embodiment or example. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples. Further, one skilled in the art can engage and combine the different embodiments or examples described in this specification.
While embodiments of the present invention have been shown and described above, it will be understood that the above embodiments are illustrative and not to be construed as limiting the invention, and that variations, modifications, alternatives and variations may be made to the above embodiments by one of ordinary skill in the art within the scope of the invention.
Claims (6)
1. A detection device for cleaning residual protein, comprising:
the sampling assembly comprises a sampling swab (1) and a transparent reagent tube (2); a reagent storage area (22) sealed by a first sealing diaphragm (21) is arranged in the reagent tube (2);
a second sealing diaphragm (23) is arranged in the reagent tube (2), and the second sealing diaphragm (23) is arranged at one side of the first sealing diaphragm (21) away from the open end of the reagent tube (2) at intervals so as to divide the reagent storage area (22) into a sealed reagent storage area A (221) and a reagent storage area B (222);
the detection part comprises a constant temperature culture block (3), a photoelectric detection assembly (4) and a controller (5); a constant temperature culture cavity (31) is arranged in the constant temperature culture block (3), and a test tube insertion hole (33) and a penetrating detection hole (32) are respectively formed in the top wall and the opposite side walls of the constant temperature culture block (3) corresponding to the constant temperature culture cavity (31); the reagent tube (2) can be inserted into the constant temperature culture cavity (31) from the test tube insertion hole (33), and the outer wall of the reagent storage area (22) corresponds to the detection hole (32); the transmitting end and the receiving end of the photoelectric detection assembly (4) are respectively correspondingly arranged at the outer side orifice positions of the two detection holes (32) so as to detect the reaction liquid in the reagent storage area (22); the photoelectric detection assembly (4) is electrically connected with the controller (5);
The sampling swab (1) comprises a swab rod (11), a sampling head (12), a sliding sealing plug (14) and a handle (13);
the sampling head (12) is fixed at one end of the swab rod (11) and the free end of the sampling head is a sample collecting end; one end of the sliding sealing plug (14) is fixed at the other end of the swab rod (11); the handle (13) is fixed at the other end of the sliding sealing plug (14); the sampling head (12) can be inserted into the tube from the open end of the reagent tube (2), and the sliding sealing plug (14) can be in sliding sealing connection with the reagent tube (2); the sampling head (12) of the sampling swab (1) can be inserted into the tube from the open end of the reagent tube (2) and sequentially pierce the first sealing diaphragm (21) and the second sealing diaphragm (23) downwards, and the sample at the sample collecting end is mixed with two detection reagents in the reagent storage area A (221) and the reagent storage area B (222) to form a reaction solution;
a wiper assembly (6); the liquid scraping component (6) consists of a plurality of liquid scraping elastic sheets (61); the plurality of scraping elastic pieces (61) are uniformly arranged along the circumferential direction of the inner wall of the reagent tube (2), are respectively positioned at the side of the first sealing diaphragm (21) far away from the second sealing diaphragm (23) and are positioned below the sampling head (12); one end of each of the plurality of liquid scraping elastic pieces (61) is fixed with the inner wall of the reagent tube (2), the other end of each of the plurality of liquid scraping elastic pieces is a free end, and the plurality of liquid scraping elastic pieces (61) are obliquely arranged downwards; the diameter of a central hole formed by the free end circumferences of the plurality of scraping elastic pieces (61) is smaller than the outer diameter of the sampling head (12);
The test tube insertion hole (33) is a guiding and positioning hole of the reagent tube (2); the bottom end of the reagent tube (2) can penetrate through the test tube insertion hole (33) so as to lead the reagent storage area (22) to be aligned with the detection hole (32);
the photoelectric detection assembly (4) comprises a light-emitting plate (41) and a receiving plate (42);
the light-emitting plate (41) and the receiving plate (42) are symmetrically fixed on two side walls of the constant temperature culture block (3); a light source (411) is fixed on the side wall of the light-emitting plate (41) close to the constant-temperature culture block (3), a photoelectric sensor (421) is arranged on the side wall of the receiving plate (42) close to the constant-temperature culture block (3), and the light source (411) and the photoelectric sensor (421) are respectively positioned at two ends in the detection hole (32); the light source (411) and the photoelectric sensor (421) are electrically connected with the controller (5);
the light source (411) is a white light source; the photoelectric sensor (421) comprises a plurality of photoelectric receiving tubes (4211), and the photoelectric receiving tubes (4211) are arranged in parallel to form an array type receiving end; the front end of each photoelectric receiving tube (4211) is provided with an optical filter (4212); the optical filters (4212) respectively correspond to different preset light transmission wave bands; the photoelectric receiving tubes (4211) are electrically connected with the controller (5) to receive and feed back the filtered optical signals of the designated wave bands.
2. The apparatus for detecting protein residues by washing according to claim 1,
the bottom of the constant temperature culture block (3) is provided with a constant temperature heating plate (34), and the side part of the constant temperature culture block (3) is provided with a temperature sensor (35) corresponding to the constant temperature culture cavity (31); the constant temperature heating plate (34) and the temperature sensor (35) are electrically connected with the controller (5).
3. The device for detecting protein residues by washing according to claim 2, wherein the reagent storage area a (221) stores sodium carbonate hydrate or not, and the reagent storage area B (222) stores copper sulfate; or copper sulfate is stored in the reagent storage area A (221), and anhydrous sodium carbonate is stored in the reagent storage area B (222).
4. A detection method of a detection device for cleaning residual protein according to any one of claims 2 to 3, comprising the steps of:
step one: wiping the surface of the cleaned detected instrument by the head of the sampling swab (1) so as to collect residual protein on the detected instrument;
step two: inserting a sample collection end of the sampling swab (1) for collecting a sample into the tube from the open end of the reagent tube (2) and downwards piercing the sealing diaphragm I (21), and enabling the head of the sampling swab (1) to enter the reagent storage area (22) so as to fully mix and contact the collected residual protein sample with the detection reagent in the reagent storage area (22) to form a reaction solution;
Step three: pulling up the sampling swab (1) and enabling a sample collection end of the sampling swab to leave the reagent storage area (22), and inserting a part of the reagent tube (2) corresponding to the reagent storage area (22) into a constant temperature culture cavity (31) for constant temperature culture of reaction liquid so as to perform a color change reaction;
step four: the controller (5) is used for controlling the emitting end of the photoelectric detection component (4) to emit optical signals, the optical signals pass through the detection hole (32) and penetrate through the reagent storage area (22) to be received by the receiving end of the photoelectric detection component (4), and the photoelectric detection component (4) is used for converting the received optical signals into electric signals to be transmitted and fed back to the controller (5);
step five: and the controller (5) processes the electric signals fed back by the photoelectric detection component (4) by adopting a weighted mobility analysis method, and finally, quantitative values of the cleaning residual proteins are obtained.
5. The method for detecting a protein residue washing apparatus according to claim 4, wherein,
the specific steps of collecting the residual protein on the detected instrument in the first step are as follows: holding the handle (13) in hand, pulling out the sampling swab (1) from the reagent tube (2), wetting the sampling head (12) of the sampling swab (1) by distilled water, manually operating the sampling head (12) to wipe the surface of the detected instrument, and collecting cleaning residues on the surface of the detected instrument;
The specific steps of the second step are as follows: reinserting the sampled sampling swab (1) into the reagent tube (2), firstly inserting the sampling head (12) from the top opening of the reagent tube (2), pushing the handle (13), sequentially puncturing the sealing diaphragm I (21) and the sealing diaphragm II (23) by using the sampling head (12), enabling the sampling head (12) to enter the reagent storage area A (221), enabling anhydrous sodium carbonate and copper sulfate reagent to be fully mixed and contacted in the reagent storage area A (221), and shaking the test tube forcefully to enable collected cleaning residues to fall off the sampling head (12) and enable the reagent storage area A (221) to be subjected to internal reference color change reaction;
the specific steps of the third step are as follows: preheating the constant temperature culture cavity (31), controlling and starting the constant temperature heating plate (34) through the controller (5), and regulating and controlling the temperature of the constant temperature culture cavity (31) to be 60 ℃ to be unchanged; inserting the reagent tube (2) with the collected cleaning residual protein into the constant temperature culture cavity (31) through a test tube inserting hole (33), enabling the bottom end of the reagent tube (2) to be abutted with the bottom end of the constant temperature culture cavity (31) so as to ensure that the reagent storage A area (221) is aligned with the detection hole (32), and carrying out constant temperature culture;
The specific steps of the fourth step are as follows: the controller (5) is used for turning on the light source (411) to emit white light to irradiate and penetrate through the test tube wall corresponding to the reagent storage area A (221), and the white light is absorbed and filtered by the reaction liquid in the reagent storage area A (221) to generate light which penetrates through the test tube wall to be output and reaches the photoelectric sensor (421); the controller (5) performs periodic data acquisition on a plurality of photoelectric receiving tubes (4211); the data collected in each period is D (i, t), wherein i=1, 2,3 … N, t=1, 2,3 … N, i is the number of the photoelectric receiving tubes (4211), N is the number of the photoelectric receiving tubes (4211), light transmission bands different from the plurality of optical filters (4212) are in one-to-one correspondence, the band numbers of the light transmission bands are Ch1, ch2, ch3 … Ch, t are sampling moments, the sequences are numbered from 1, and N is the moment number corresponding to the last sampling value;
the specific steps of the fifth step are as follows: firstly, carrying out weighted normalization on the sample value of each photoelectric receiving tube (4211); determining each weighting value omega based on the gain of the photo-receiving tube 4211 and the attenuation value of the optical filter 4212 i I=1, 2,3 … n; each sample value is multiplied by a corresponding weighting value to obtain a normalized sample value:
σ i （t）=ω i x D (i, t), where i=1, 2,3 … N, t=1, 2,3 … N;
secondly, normalizing the Bai Guangguang source intensity; in the case of initially inserting the reagent tube (2) and turning on the white light source, the data of each photoelectric receiving tube (4211) is sampled once, and the weighted sampling value in the case of initially inserting the reagent tube (2) is represented by t=0, the normalized coefficient λ of the white light source intensity is represented as:
；
then, calculating the band mobility of each photoelectric receiving tube (4211); band mobility S i The calculation formula of (2) is as follows:
，i=1，2，3…n-1；
d i is the distance between the center wavelength of the light transmission wave band corresponding to the photoelectric receiving tube (4211) with the number i and the center wavelength of the light transmission wave band with the wave band number Chn; band mobility is the full path delta of each transmission band relative to the transmission band with band number Chn since the start of incubation;
finally, the band mobilities of all the photo-receiving tubes (4211) are summed up and the current instant residual protein reading R (n) can be obtained taking into account the normalized coefficient of the white light source intensity, expressed as:
。
6. the method for detecting a detecting device for washing residual protein according to claim 5, wherein an alarm threshold is set for R (n) as needed; at any time of incubation, when the value of R (n) exceeds the alarm threshold, indicating that the residual protein reading has exceeded the expected value, the early end of the incubation process can be controlled; when the cultivation is finished, the controller (5) displays the reading value of the detection device; the detection curve of the mapping relation between the reading of the detection device and the residual protein can be obtained by preparing the tested object with known protein quantity; the reading value of the detection device can be converted into the amount of the washing residual protein in μg/ml according to the detection curve every time a sample of unknown residual protein amount is tested.
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