CN117647641A - Combined detection kit for iron ions, active oxygen and nitrite - Google Patents
Combined detection kit for iron ions, active oxygen and nitrite Download PDFInfo
- Publication number
- CN117647641A CN117647641A CN202311624350.0A CN202311624350A CN117647641A CN 117647641 A CN117647641 A CN 117647641A CN 202311624350 A CN202311624350 A CN 202311624350A CN 117647641 A CN117647641 A CN 117647641A
- Authority
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- China
- Prior art keywords
- detection
- active oxygen
- nitrite
- iron ions
- kit
- Prior art date
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- Pending
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- 238000001514 detection method Methods 0.000 title claims abstract description 139
- -1 iron ions Chemical class 0.000 title claims abstract description 65
- XEEYBQQBJWHFJM-UHFFFAOYSA-N iron Substances [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 title claims abstract description 44
- 239000001301 oxygen Substances 0.000 title claims abstract description 44
- 229910052760 oxygen Inorganic materials 0.000 title claims abstract description 44
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 title claims abstract description 43
- IOVCWXUNBOPUCH-UHFFFAOYSA-M Nitrite anion Chemical compound [O-]N=O IOVCWXUNBOPUCH-UHFFFAOYSA-M 0.000 title claims abstract description 42
- 229910052742 iron Inorganic materials 0.000 title claims abstract description 42
- 238000005070 sampling Methods 0.000 claims abstract description 74
- 238000012360 testing method Methods 0.000 claims abstract description 66
- NLKNQRATVPKPDG-UHFFFAOYSA-M potassium iodide Chemical compound [K+].[I-] NLKNQRATVPKPDG-UHFFFAOYSA-M 0.000 claims abstract description 36
- 239000007788 liquid Substances 0.000 claims abstract description 19
- 238000003908 quality control method Methods 0.000 claims abstract description 19
- 229920002472 Starch Polymers 0.000 claims abstract description 15
- 239000008107 starch Substances 0.000 claims abstract description 15
- 235000019698 starch Nutrition 0.000 claims abstract description 15
- 239000003153 chemical reaction reagent Substances 0.000 claims abstract description 14
- 238000000605 extraction Methods 0.000 claims abstract description 14
- 229920000259 polyoxyethylene lauryl ether Polymers 0.000 claims abstract description 14
- 239000000276 potassium ferrocyanide Substances 0.000 claims abstract description 14
- XOGGUFAVLNCTRS-UHFFFAOYSA-N tetrapotassium;iron(2+);hexacyanide Chemical compound [K+].[K+].[K+].[K+].[Fe+2].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-] XOGGUFAVLNCTRS-UHFFFAOYSA-N 0.000 claims abstract description 14
- AKHNMLFCWUSKQB-UHFFFAOYSA-L sodium thiosulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=S AKHNMLFCWUSKQB-UHFFFAOYSA-L 0.000 claims abstract description 11
- 235000019345 sodium thiosulphate Nutrition 0.000 claims abstract description 11
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims abstract description 10
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- 238000000034 method Methods 0.000 claims description 46
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- DBMJMQXJHONAFJ-UHFFFAOYSA-M Sodium laurylsulphate Chemical compound [Na+].CCCCCCCCCCCCOS([O-])(=O)=O DBMJMQXJHONAFJ-UHFFFAOYSA-M 0.000 claims description 9
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- 235000017281 sodium acetate Nutrition 0.000 claims description 8
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Classifications
-
- 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
- G01N33/52—Use of compounds or compositions for colorimetric, spectrophotometric or fluorometric investigation, e.g. use of reagent paper and including single- and multilayer analytical elements
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
- A61B10/00—Other methods or instruments for diagnosis, e.g. instruments for taking a cell sample, for biopsy, for vaccination diagnosis; Sex determination; Ovulation-period determination; Throat striking implements
- A61B10/0045—Devices for taking samples of body liquids
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
- A61B10/00—Other methods or instruments for diagnosis, e.g. instruments for taking a cell sample, for biopsy, for vaccination diagnosis; Sex determination; Ovulation-period determination; Throat striking implements
- A61B10/0045—Devices for taking samples of body liquids
- A61B10/0051—Devices for taking samples of body liquids for taking saliva or sputum samples
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/40—Concentrating samples
- G01N1/4055—Concentrating samples by solubility techniques
-
- 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
- G01N21/01—Arrangements or apparatus for facilitating the optical investigation
-
- 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
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/40—Concentrating samples
- G01N1/4055—Concentrating samples by solubility techniques
- G01N2001/4061—Solvent extraction
-
- 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
- G01N21/01—Arrangements or apparatus for facilitating the optical investigation
- G01N2021/0106—General arrangement of respective parts
- G01N2021/0112—Apparatus in one mechanical, optical or electronic block
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Hematology (AREA)
- Physics & Mathematics (AREA)
- Biomedical Technology (AREA)
- Analytical Chemistry (AREA)
- Molecular Biology (AREA)
- Biochemistry (AREA)
- General Physics & Mathematics (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Surgery (AREA)
- Medical Informatics (AREA)
- Heart & Thoracic Surgery (AREA)
- Urology & Nephrology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Plasma & Fusion (AREA)
- Biotechnology (AREA)
- Cell Biology (AREA)
- Microbiology (AREA)
- Pulmonology (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention discloses a combined detection kit for iron ions, active oxygen and nitrite, which relates to the technical field of biochemistry and comprises a sampler, a sampling tube and a detection plate, wherein a sample extraction liquid is arranged in the sampling tube, a test strip is arranged in the detection plate, a detection indicator, a quality control indicator and a middle line reagent are arranged on the test strip in a drying way, wherein the detection indicator comprises starch, potassium ferrocyanide, polyoxyethylene lauryl ether, potassium iodide, sodium thiosulfate, calcium chloride and an alkaline pH regulator.
Description
Technical Field
The invention relates to the technical field of biochemistry, in particular to a combined detection kit for iron ions, active oxygen and nitrite.
Background
Iron ions (Fe) 3+ ) Reactive oxygen species (Reactive Oxygen Species/ROS, oxygen radicals) and Nitrite (NO) 2 - ) The lung inflammation airway mucus is a body fluid and gel protective layer existing on the inner surface of the airway, and mainly consists of airway mucin, moisture, inorganic salt and the like. Pulmonary inflammation (pneumonia) is a group of inflammation occurring in the terminal airways, alveoli and pulmonary parenchyma in the lung, and can be caused by pathogenic microorganism infection, physical and chemical factors, immune injury, allergy and other factors, and has great threat to life health, but the early stage is caused by downward spreading of upper respiratory tract infection, and has no additional characteristic symptoms, chest X-ray/CT images are usually not parallel to the pneumonia, the images can be changed abnormally, the pneumonia can be formed without obvious X-ray image expression, and peripheral blood leucocytes can also be normal, so that diagnosis and treatment delay is easily caused. Taking a novel coronal (new coronal) virus infection as an example, the new coronal infection is mostly manifested by symptoms of upper respiratory tract infection such as fever, cough, headache, general muscular soreness, hypodynamia and the like, most patients can lighten the symptoms after one week and enter a recovery period, and can be in home rest, but partial patients can possibly generate pneumonia, and the patients develop diffuse lung tissue injury including extensive exudation and hemorrhage, the destruction of alveolar epithelium and reactive hyperplasia, ventilation dysfunction and dyspnea, if missed and cured in time, death can be caused, and the development is convenient and easy to use by the rapid detection of airway inflammation markers such as iron ions, active oxygen, nitrite and the like, and the development is convenient and easy to use by the lung inflammation screening, tracking and monitoring method and product suitable for popularization, so that the prevention, control, diagnosis and treatment of respiratory tract infection and pneumonia can be greatly improved and promoted.
The existing iron ion detection method mainly comprises a colorimetry and a titration method, and the existing active oxygen detection method mainly comprises a chemiluminescence method, a fluorescent probe, a spectrophotometry method, an electron paramagnetic resonance method and the like. The existing nitrite or nitric oxide detection methods mainly comprise an absorbance method (colorimetry), a fluorescent probe method, a chemiluminescent method (luminol method), a sensor method (electrochemistry/optics) and the like, and the methods need to carry out sample processing steps with strong speciality, cannot be used for instant quick detection (POCT), cannot carry out multi-index synchronous joint detection, and are not beneficial to popularization and application such as screening and self-detection;
the system and the method for qualitative and convenient detection of hydrogen peroxide and nitrite are provided in the Chinese patent application 202211539935.8 of the invention, namely an airway hydrogen peroxide detection method and a kit, and the Chinese patent application 202211545905.8 of the invention, namely an airway nitric oxide rapid detection method and a detection kit, respectively, but the system and the method still have the defects that part of people, particularly children, are difficult to expect samples, part of people samples are too viscous and difficult to process, and the sampling and processing processes are easy to be interfered, are unfavorable for general application, meanwhile, the production process of the system and the color development definition of the detection result also need to be further enhanced and improved, and the system needs to be matched and applied to the combined detection containing iron ions so as to be favorable for application in early-stage screening detection and tracking monitoring of pneumonia.
Disclosure of Invention
In summary, a rapid, simple and convenient method and product for detecting iron ions, active oxygen and nitrite in a combined manner are lacking at present, and therefore, the invention provides the method and the kit for detecting iron ions, active oxygen and nitrite in a combined manner, which are safe, noninvasive, convenient, easy to use, reliable in performance and capable of being used for timely detection, and can be used in non-professional scenes such as home or communities, so as to facilitate screening and self-detection, and solve the problems in the background art.
In order to achieve the above purpose, the present invention provides the following technical solutions: a combined detection method of iron ions, active oxygen and nitrite comprises the following steps:
s1: sampling, namely sampling the oral cavity by using a throat swab sampler, wherein for a sampling part which can be in direct contact, a user can directly wipe the sampling part by using the throat swab sampler to sample; for airway mucus detection, after a user washes mouth and cleans the mouth, the user drinks a mouthful of water, beats the chest or back 10 times, deeply breathes and forcefully cough 6 times, so that a small amount of airway mucus is discharged from the airway and accumulated in the mouth, then the user inserts the throat swab sampler into the upper rear end of the mouth and tongue, and the mouth is closed for sampling for about 3-5 seconds;
s2: adding a sample, immersing the sampling end of the sampled sampler into a sample extract containing oxalic acid, sodium chloride and 8-hydroxyquinoline, and mixing;
s3, pipetting, namely transferring the mixed sample extract to a test strip, wherein a detection indicator, a quality control indicator and an intermediate line reagent are arranged on the test strip in a drying manner, and the detection indicator comprises the components of starch, potassium ferrocyanide, polyoxyethylene lauryl ether, potassium iodide, sodium thiosulfate, calcium chloride and an alkaline pH regulator;
s4, performing S4; observing, standing until the quality control indicator changes color to display mauve, observing the color of the detection indicator, and if the detection indicator shows that the blue-black to black-brown color is a positive result, the content of iron ions, active oxygen and/or nitrite reaches or exceeds a detection threshold value; and if the detection indicator does not develop color or has a light color as a negative result, the content of the iron ions, the active oxygen and/or the nitrite is lower than the detection threshold value.
Preferably, the detection indicator comprises the components of 2.0-8.0% (w/v) starch, 100-800mM potassium ferrocyanide, 1.0-7.0M potassium iodide, 50-300mM calcium chloride, 0.5-3.0mM pH regulator, 1-20mM sodium thiosulfate and 0.5-2.0% (w/v) polyoxyethylene lauryl ether.
Preferably, the composition of the sample extract comprises 50-500mM oxalic acid, 50-300mM sodium chloride and 0.5-2mM 8-hydroxyquinoline.
Preferably, the composition of the alkaline pH regulator comprises one or more of sodium acetate, sodium bicarbonate, tris, sodium carbonate and sodium hydroxide.
Preferably, the composition of the quality control indicator comprises 5-30mM neutral red and 10-60mM alkaline pH regulator.
Preferably, the test strip is marked with an S area for sample adding at the end part, a T area for detection is marked at the position 12-18mm away from the S area, the detection indicator is arranged in the T area, an R area is marked at the position 4-6mm away from the T area, the middle line reagent is arranged in the R area, a C area is marked at the position 6-9mm away from the R area, and the quality control indicator is arranged in the C area.
Preferably, the composition of the intermediate line reagent comprises 5% -15% (w/v) polyoxyethylene lauryl ether and 5% -15% (w/v) sodium dodecyl sulfate.
Preferably, the throat swab sampler is a disposable sterile sampling swab for detecting new crown nucleic acid, a swab handle material is made of hollow Polystyrene (PS), polypropylene (PP) or ABS plastic, and a sampling end sampling swab head is made of PE synthetic fibers, polyester fibers, polypropylene fibers, artificial fibers and other synthetic fibers; the primary sampling amount is about 100 mg.+ -.15 mg or 100. Mu.l.+ -.15. Mu.l.
Preferably, the drying process of the test strip is as follows: and (3) the temperature is kept for 5 to 20 minutes in a 50+/-10 ℃ oven or an air circulation incubator in a dark place.
In addition, the invention also provides a kit using the combined detection method of the iron ions, the active oxygen and the nitrite.
The combined detection kit for the iron ions, the active oxygen and the nitrite comprises a throat swab sampler, a sampling tube and a detection plate, wherein a sample extraction liquid is arranged in the sampling tube, and a test strip is arranged in the detection plate.
Preferably, the test strip in the test plate is a filter paper strip 68mm×6mm×0.7mM (length×width×thickness), and 3 μl of the detection color-developing agent is dried at a distance of 27mM from the right end of the filter paper strip, and the composition is 2% (w/v) starch, 1% (w/v) polyoxyethylene lauryl ether, 750mM potassium ferrocyanide, 2M potassium iodide, 250mM calcium chloride, 15mM sodium thiosulfate, 2.5mM sodium acetate, 10% (w/v) polyoxyethylene lauryl ether, 10% sodium dodecyl sulfate, and 3 μl of the quality control color-developing agent is dried at a distance of 38mM from the right end of the filter paper strip, and the composition is 15mM neutral red, 45mM sodium bicarbonate.
Preferably, the sample extract in the sample tube in the kit has a volume of 300. Mu.l/serving and a composition of 200mM oxalic acid, 100mM sodium chloride, 2mM 8-hydroxyquinoline.
Preferably, the sampling tube in the kit is a tube body and a tube cover separated, and the tube cover can be unscrewed or screwed on according to the requirement.
Preferably, the detection plate adopts a general colloid Jin Kake, and is made of Polypropylene Plastic (PP).
Preferably, the concentrations of iron ions, active oxygen and nitrite ions detected are not less than 1mM, respectively.
The principle of the invention is based on the suction filtration chromogenic qualitative determination of potassium ferrocyanide combined with ferric ions, active oxygen and nitrite, wherein iodine ions (I-) are oxidized under acidic conditions to generate iodine molecules (I2) combined with starch. After a user washes mouth and cleans the mouth, the lung airway accelerates by drinking water, beating the front chest or back and making deep breathing and forceful deep cough, part of airway mucus carrying lung secretion is discharged into the mouth, then a throat swab sampler is utilized for sampling and detecting, if inflammation and tissue injury local hemorrhage exist in a sample, iron ions (usually existing in iron-containing flavins formed by decomposing hemoglobin), active oxygen (mainly accumulated in the form of hydrogen peroxide) or nitrite are accumulated and raised in the airway mucus.
In the sample extract provided by the invention, oxalic acid can crack a sample, denature and precipitate protein, and dissolve and release iron ions, active oxygen and nitrite; oxalic acid and sodium chloride provide the acidic pH and ionic conditions required to oxidize iodide ions. After a sample is mixed with a sample extraction liquid to form a sample solution and is dripped on a test strip, the sample solution diffuses to wet and dissolve a detection indicator in a T area, and iodine ions in the detection indicator are rapidly oxidized to generate iodine molecules and immediately combined with starch for color development. In the formula of the detection color developing agent provided by the invention, 2.0% -8.0% (w/v) starch and 1.0-7.0M potassium iodide provide an oxidation color developing base system, and ratios outside the range all lead to obvious reduction of color developing definition; the addition of 0.5% -2.0% (w/v) polyoxyethylene lauryl ether and 100-800mM potassium ferrocyanide can promote the dissolution of starch mother liquor and the drying and fixation of detection indicators in the test strip T area in the production and preparation process, and the local high-concentration polyoxyethylene lauryl ether can delay the diffusion of the starch solution, so that the clear aggregation of color development is enhanced; the potassium ferrocyanide can react with iron ions in the sample to generate precipitate, so that the clear readability of color development is further enhanced; the potassium ferrocyanide 100-800mM, the sodium thiosulfate 1-20mM and the alkaline pH regulator 0.5-3.0mM can eliminate iodine molecular background possibly existing in the potassium iodide test solution, inhibit slow oxidation of dissolved oxygen and oxygen in air to iodide ions, and enable the test strip to be preserved for a long time under the condition of normal temperature drying.
In summary, the invention has the beneficial effects that:
1. according to the invention, through a suction filtration color reaction system, three indexes of inflammation such as iron ions, active oxygen and nitrite are synchronously detected, a user can qualitatively judge whether the 3 indexes of inflammation are remarkably abnormal in a few minutes through simple and continuous operation, and the method is particularly expected to be used for POCT self-test rapid detection and noninvasive early screening of pulmonary inflammation;
2. the invention can detect by using the sampling detection method of the invention before the symptom is obvious or expectoration is formed, the process is safe, convenient and harmless, and does not need invasive means, and simultaneously, the invention can detect pneumonia characterized by pneumonia and lung injury hemorrhage or airway nitric oxide elevation caused by the rising of active oxygen or inactive oxygen of the pneumonia in the stage of obvious injury hemorrhage, and can achieve better pneumonia detection sensitivity than the detection of only detecting a certain single index;
3. compared with the traditional exhaled breath detection, the three detected target substances lack of volatility and mainly exist in airway mucus: iron ions exist in the ferrioxacin, the half-life period of nitric oxide gas is very short, the nitric oxide gas mainly exists in the form of nitrite, and active oxygen in the lung mainly exists in the form of lipid peroxide and hydrogen peroxide, so that the invention has better applicability and sensitivity than the detection of exhaled air. Meanwhile, compared with the conventional exhaled air sampling, the airway mucus sampling can eliminate air flow interference, has better anti-interference performance, is easier to use and can achieve better detection efficiency;
4. compared with the conventional colorimetric method or fluorescent method, the method has the advantages that quantitative integrated operation is not needed in the use process, additional equipment is not needed, background interference is eliminated from the color development threshold value of the detection area, and therefore, the method is beneficial to qualitative and visual judgment, is simpler, faster and more suitable for popularization;
5. compared with various blood detection methods, the method has the advantages that the lung inflammation condition is directly detected and prompted, the sensitivity and the specificity are better than those of blood detection, blood is not required to be drawn, and the method has excellent safety and noninvasive characteristics;
6. compared with the traditional sputum detection method, the method can collect and detect airway mucus samples in a specified mode in a conventional period, and has better sensitivity and applicability;
7. the invention has the advantages of safety, no wound, convenience, easy use, reliable performance, moderate price and the like, is favorable for popularization and application, is expected to solve the problem of insufficient convenience and easy use of inflammation detection, and when the invention is used for pulmonary inflammation related tests, the positive detection result prompts that the lung of a subject possibly has lung tissue injury hemorrhage or neutrophilic granulocyte inflammation/inflammation storm or eosinophilic granulocyte inflammation, and the subject should go to a professional medical institution for related examination and diagnosis. The invention is applied to early screening and dynamic monitoring of lung inflammation of respiratory tract infection patients and high risk groups, and is greatly beneficial to improving and promoting prevention, control, diagnosis and treatment of lung inflammation.
Drawings
In order to more clearly illustrate the embodiments of the invention or the technical solutions in the prior art, the drawings that are used in the description of the embodiments or the prior art will be briefly described, it being obvious that the drawings in the description below are only some embodiments of the invention, and that other drawings can be obtained from these drawings without inventive effort for a person skilled in the art.
FIG. 1 is a schematic diagram of test strips and detection results in a combined detection kit for iron ions, active oxygen and nitrite in the invention;
FIG. 2 is a photograph of a finished test strip and detected fruits in a combined detection kit for iron ions, active oxygen and nitrite according to the present invention;
FIG. 3 is a schematic diagram of the composition structure of a reagent box in the combined detection kit for iron ions, active oxygen and nitrite in the invention;
FIG. 4 is a front view of a sample tube containing a sample and sample extract mixture in a combined detection kit for iron ions, active oxygen and nitrite according to the present invention;
FIG. 5 is a photograph of the finished product of the reagent kit and the detected fruits in the combined detection kit for iron ions, active oxygen and nitrite.
The index marks in the drawings are as follows: 320. the test strip sample adding area; 321. detecting an indication area (T area); 322. a middle line region (R region); 323. a quality control indication region (region C); KA. A throat swab sampler; KB. A sampling tube for containing sample extract; KC. A detection plate with a test strip inside; KA1, sampling end of sampler; KB1, sampling tube body; KB2, tube cap; KB3, sample extract; KC1, detecting a card shell; KC2, detection zone (T); KC3, a quality control region (C); KC4, well (S); KC5, clamping groove
Detailed Description
All of the features disclosed in this specification, or all of the steps in a method or process disclosed, may be combined in any combination, except for mutually exclusive features and/or steps.
Any feature disclosed in this specification (including any accompanying claims, abstract and drawings), may be replaced by alternative features serving the same, equivalent or similar purpose, unless expressly stated otherwise. That is, each feature is one example only of a generic series of equivalent or similar features, unless expressly stated otherwise.
The present invention will now be described in further detail with reference to the following specific embodiments of fig. 1-5, it being understood that the specific embodiments are merely illustrative of the invention and not limiting of the invention, and that the embodiments described herein are merely some, but not all,
referring to fig. 1-5, an embodiment of the present invention is provided: the position 13mm away from the bottom of the test strip is set as a sample adding area (S area) 320, and the sample solution to be detected can be dripped or immersed into the area; 12mM above the S region is a detection indication region (T region) 321, and 2% (w/v) of starch, 750mM potassium ferrocyanide, 1% (w/v) of polyoxyethylene lauryl ether, 250mM of calcium chloride, 2M of potassium iodide, 15mM of sodium thiosulfate and 2.5mM of sodium acetate are added dropwise; a middle line indication area (R area) 322 is arranged 5mm above the T, and 10% (w/v) sodium dodecyl sulfate and 10% (w/v) polyoxyethylene lauryl ether are added dropwise; a quality control indication region (C region) 323 is arranged 5mM above the R region, and 15mM neutral red and 45mM sodium bicarbonate are added dropwise. After drying treatment for 5 minutes at 50 ℃, the test strip can be directly used for sample detection, can also be sealed, dried and stored at normal temperature for standby, and is observed to detect an indication area (T), wherein the color change shows that the bluish black to blackish brown color is a positive result, the content of iron ions, active oxygen and/or nitrite reaches or exceeds a detection threshold value, no color development or light color is a negative result, and the content of iron ions, active oxygen and/or nitrite is lower than the detection threshold value;
the kit mainly comprises a throat swab sampler KA, a built-in sample extraction liquid sampling tube KB and a detection plate KC with a built-in test strip, wherein the detection plate KC1 is colloid Jin Kake, the appearance specification is 70X 20mm, the specification of a sample adding hole is 6X 3mm, and the specification of an interpretation window is 16X 3.5mm. A clamping groove KC5 for accommodating a test strip is formed in the test strip, the specification is 60 multiplied by 4mm, the test strip is fixedly arranged in the clamping groove KC5, the size of the test strip is determined according to the size of the indication groove, and a sample control line can be omitted;
the sample extraction liquid is arranged in the sampling tube KB, the volume of the sample extraction liquid is 300 mul, the sample extraction liquid consists of 100mM sodium chloride, 200mM oxalic acid and 2mM 8-hydroxyquinoline, the sampled throat swab KA is immersed in the sample extraction liquid, and the sample extraction liquid is mixed by shaking the nut tube;
the specific implementation process comprises the following steps: after sampling by a throat swab sampler, immersing a sampling end into a sampling tube with a built-in sample extraction liquid, breaking along a notch, retaining the sampling end in the tube, sealing the sampling tube, severely shaking for about 5 seconds, enabling the sampling end to be completely immersed into the sample extraction liquid, dripping liquid in the tube into a sample adding hole (S) of a detection plate, standing until a quality control area (C) changes color to display mauve, observing a detection area (T), changing the color to display blue-black to black-brown to be a positive result, and indicating that the content of iron ions, active oxygen or nitrite in the sample reaches or exceeds a detection threshold value.
The invention also provides a comparison experiment example of throat swab sampling and expectoration sampling test
We found in previous experimental studies that: after the user washes mouth and cleans the oral cavity, the user drinks a mouthful of water, beats the chest or back 10 times, breathes deeply, and cough forcefully for 6 times, so that a small amount of airway mucus is discharged from the airway and is gathered into the oral cavity, and then the liquid in the oral cavity is expectorated, collected and detected, so that mucin MUC5AC or mRNA thereof specifically expressed by the airway can be detected at 100%, and airway mucus samples can be collected. However, in the expanded popularization, it is found that a part of people, especially the young children, are difficult to expectorate the oral liquid, so that sampling fails, and in order to ensure the sampling success rate, the comparison and verification are carried out by adopting a throat swab sampling mode and an autonomous expectoration sampling mode of a 202211539935.8 (or 202211545905.8) method respectively. Throat swab sampling mode: after the user has followed the foregoing operation to cause a small amount of airway mucus to drain from the airway and accumulate in the mouth, the user need not expectorate the collection, but instead wipes the mouth with a throat swab sampler, dips the sampling end of the sampler into the sample extract, breaks along the score, and leaves both the sample and sampling end in the extract to complete the sampling. Autonomous expectoration sampling mode: the user expectorates the collection after following the foregoing procedure to encourage a small amount of airway mucus to drain from the airway and accumulate in the mouth. We recruited 30 healthy volunteers, including 20 adults and 10 infants between 1-5 years of age, for two sampling patterns comparison of different periods of time, 1-time daily, 3 days in succession, wherein the adults were all-process self-sampling (pharyngeal swab sampling or 202211539935.8 expectoration sampling of the present invention), and the infants were sampled by parents using pharyngeal swab after 6 coughs were instructed by parents. After sampling, detecting the weight of the sampling tube by comparing the weights of the sampling tube before and after sampling, and calculating to obtain the sampling weight; and the sampling liquid-transfering device is used for measuring the sampling volume, so that the throat swab sampling mode provided by the invention can successfully sample adult or infant volunteers, and the sampling fluctuation is smaller: the primary sampling amount is about 100+/-15 mu l (volume) or 100+/-15 mg (weight), and the sputum-expectoration sampling mode of 202211539935.8 (or 202211545905.8) is successful in the morning and evening of adults, but the sampling amount is about 850+/-185 mu l (volume) or 850+/-150 mg (weight), and 5 adult sampling in the afternoon is unsuccessful; the infants had 3 sampling failures at each period, and the specific statistics are shown in Table 1 below.
TABLE 1 statistics of test of throat swab and expectoration samples of volunteers
The invention provides an experimental example for analyzing specific components of a test strip
The test strips were 68mm by 6mm long by 6mm wide filter strips. A detection area (T) is marked at a position 27mm away from the bottom end of the filter paper strip, and a strip of mixed solution of starch, potassium ferrocyanide, polyoxyethylene lauryl ether (Brij-35), calcium chloride, potassium iodide, sodium thiosulfate and sodium acetate is uniformly dripped into the pipette tip; a mixed solution strip of Sodium Dodecyl Sulfate (SDS) and polyoxyethylene lauryl ether (Brij-35) is uniformly dripped into a middle line region (R) marked 5mm above the mixed solution strip by a pipette tip; a quality control area (C) is marked 5mm above the liquid transfer device, and a neutral red and alkaline pH regulator solution strip is evenly dripped into the liquid transfer device; the reagent formulations of the T region, the R region and the C region are shown in Table 1, and the volumes of the reagent formulations are 3 mu L. The composition of the sample extract was: 100mM sodium chloride, 200mM oxalic acid, 8-hydroxyquinoline solution. The negative control was purified water, and the 3 positive controls were 1mM H2O2, 1mM FeCl3, 1mM NaNO2, respectively. The detection method is carried out according to instructions. After the operation is finished, obvious mauve color can be observed at the C position, and the correct operation is indicated; if 3 positive controls are observed to change color in the T region, the positive results are shown from blue black to black brown. Therefore, the test strips prepared in the concentration range of the tested reagent can reach the expected detection result (Table 2).
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Table 2 control test and statistics of test strip composition
The invention provides an experimental example of the analysis of specific components of a sample extract
Test strip specification and preparation see description text and example 2. The T region is dripped with 750mM potassium ferrocyanide, 2% (w/v) starch, 2M potassium iodide, 15mM sodium thiosulfate, 250mM calcium chloride, 1% (w/v) Brij-35, 2.5mM sodium acetate solution; dropwise adding 10% (w/v) SDS and 10% (w/v) Brij-35 solution into the R region; 15mM neutral red and 45mM sodium bicarbonate were added dropwise to the C region. 3 μl of each of the T region and the C region of the test strip was added dropwise. The sample extract formulations are shown in table 3. The negative control was purified water, and the 3 positive controls were 1mM H2O2, 1mM FeCl3, 1mM NaNO2, respectively. The detection method is carried out according to instructions. After the operation is finished, the C area can be observed to be obviously purple red, and the correct operation is indicated; no obvious color development is observed in the negative control, and the detection result is negative; obvious blue-black to black-brown color can be observed for all the 3 positive control substances, and the positive detection result is indicated. Therefore, the sample extract prepared in the concentration range of the tested reagent can reach the expected detection result.
TABLE 3 composition of sample extracts
The invention provides an experimental example of the influence of the residual common beverage and medicine in the sample on the judgment of the result
Test strip specification and preparation see description text and example 2. The T region is dripped with 750mM potassium ferrocyanide, 2% (w/v) starch, 2M potassium iodide, 15mM sodium thiosulfate, 250mM calcium chloride, 1% (w/v) Brij-35, 2.5mM sodium acetate solution; the R area is dripped with 10% (w/v) Brij-35 and 10% (w/v) SDS; 15mM neutral red and 45mM sodium bicarbonate were added dropwise to the C region. 3 μl of each of the T region, R region and C region of the test strip was added dropwise. The negative control was purified water, and the 3 positive controls were 1mM H2O2, 1mM FeCl3, 1mM NaNO2. The detection method is carried out according to instructions. The interfering substances tested are shown in Table 4. The content of the interfering substance in the sample is unified to be that the conventional dosage of the substance accounts for 2% of the volume of the sample, which is higher than the possible volume ratio of the residual quantity of the substance after the conventional use and the gargling in the sampling. 3 replicates were performed for each sample, for a total of 3 runs. The result observation statistics show that after the operation is finished, the C region can observe obvious mauve, and the operation is correct; no obvious color development is observed in the negative control, and the detection result is negative; obvious blue-black to black-brown color can be observed for all the 3 positive control substances, and the positive detection result is indicated. Thus, within the concentration range tested, the tested interfering substances do not affect the detection result, i.e. do not interfere with the detection result.
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TABLE 4 interfering substances
The invention also provides a method for producing the kit
In this example, 1500 sets of planned kit production were performed. First, a lot number and a production schedule were prepared (table 5), and material preparation was performed (table 6).
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Table 5.1500 set of schedule component tables
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TABLE 6 bill of materials
Key procedure-test strip preparation: a common qualitative filter paper cut to a specification of 300mM multiplied by 68mM is marked along a line by a pencil mark as a detection area from the bottom by about 27mM, a line by a pencil mark is marked along a middle line area above the common qualitative filter paper by 5mM, a line by a pencil mark is marked along a quality control area above the common qualitative filter paper by 5mM, 3 mu l of 2% (w/v) starch, 2M potassium iodide, 750mM potassium ferrocyanide, 250mM calcium chloride, 1% (w/v) Brij-35, 15mM sodium thiosulfate and 2.5mM sodium acetate mixed solution are dripped into the detection area by a sample pen mark according to the text method of the specification, 3 mu l of 10% (w/v) Brij-35 and 10% (w/v) SDS solution are dripped into the middle line area by the mark, and 3 mu l of 60mM copper sulfate is dripped into the quality control area by the sample pen mark. After drying at 50℃for 5 minutes. After the test strip is prepared, performing spot check of key working procedures to determine that the test strip quality is qualified.
Preparing and subpackaging a sample extract: preparing 450ml according to the formula of the sample extract of the kit shown in the text of the specification, subpackaging into sampling tubes according to 300 mu l/part, screwing up tube covers, placing the sampling tubes into self-sealing bags with drying agents, sealing, temporarily storing at room temperature to obtain semi-finished products, and carrying out external packaging after the sampling inspection is qualified.
Detecting plate assembly: the test paper strip is placed in the clamping plate, the lower end of the test paper strip can be just close to the clamping groove, the upper clamping shell is fixed on the lower clamping shell, the C position and the T position respectively correspond to the quality control area and the detection area of the test paper strip, and after the workshop inspector checks and assembles qualified on site, a qualified mark is attached to the back of the detection plate. And (3) placing the detection plate into a self-sealing bag with a drying agent, sealing, temporarily storing at room temperature to obtain a semi-finished product, and carrying out external packing after the sampling inspection is qualified.
And (3) external packing: and (5) filling the semi-finished products and specifications which are subjected to the sampling inspection into an external packaging box, attaching a qualification certificate, and warehousing for later use.
The invention also provides quality inspection related to the kit
1. Key procedure spot check
And performing spot check on key procedures after the test strip is prepared, so as to determine that the test strip quality is qualified.
(1) Appearance of
The appearance size of the test strip is in accordance with the setting.
(2) Negative reference compliance
And 5 enterprises negative reference products are taken for detection, and the detection results are negative.
(3) Compliance rate of positive reference
And 5 enterprise positive reference products are taken for detection, and the detection results are positive.
(4) Detection limit
And (3) taking the enterprise detection limit reference to carry out repeated detection for 20 times, wherein the detection result of at least 19 times is positive.
2. Inspection of semifinished products
(1) Appearance of
The components are complete, the aluminum foil bag and the drying agent are well packaged, no leakage, no damage and no moisture regain are caused; the label writing is clear, the top of the indication window is stuck with a qualified mark, and the position of the window is matched with the corresponding area on the test strip.
(2) Negative reference compliance
And directly adopting a key procedure spot check result.
(3) Compliance rate of positive reference
And directly adopting a key procedure spot check result.
(4) Detection limit
And directly adopting a key procedure spot check result.
(5) Repeatability of
And (3) taking enterprise repetitive reference, and repeatedly detecting for 10 times, wherein the detection results are positive and the color development is uniform.
(6) Sample processing efficiency
Sampling according to the airway mucus sampling method disclosed by the invention, respectively detecting 5 negative samples obtained by mixing and sub-packaging airway mucus samples of 10 healthy volunteers and 5 positive samples obtained by mixing and sub-packaging airway mucus samples of 10 pneumonia patients, wherein the negative sample detection results are negative, and the positive sample detection results are positive.
3. Inspection of finished products
(1) Appearance of
The components are complete, the aluminum foil bag and the drying agent are well packaged, no leakage, no damage and no moisture regain are caused; the label writing is clear, the top of the indication window is stuck with a qualified mark, and the position of the window is matched with the corresponding area on the test strip. The total weight of the finished product should be more than or equal to 90g, which indicates that the sealed reagent is sufficient.
(2) Performance of
After the appearance check is correct, the performance indexes of the key process spot check and the semi-finished product check are directly adopted.
The invention also provides an analysis example of the use test condition of the finished product of the kit
80 healthy volunteers are collected on site in a park, the age is 20-50 years, the occupation is practitioner in the high-tech industry, the proportion of men and women is about 50%, the patients take comprehensive physical examination in one year, the patients have no abnormal condition, the patients have no medical history in the severe smoking or severe pollution industry, the patients have no lung disease history such as lung cancer or pneumonia, and the patients have no related disease family history. Some females are pregnant or lactating and feel well on their own. The staff hosts the registration, dispenses the finished product of the kit, and carries out operation explanation demonstration. The test operation was then performed by the volunteer as described in the text of the present specification, and the result was reported to the staff. And after comparison and confirmation, carrying out result statistical analysis on the staff. The result shows that the color of the region C of the kit is changed to be purple red, and the operation accuracy is 100%; no obvious color development exists in the T areas of 80 cases, and the T areas are negative, so that the pneumonia incidence risk of the tested volunteers is low.
The invention also provides an analysis example of the use test condition of the patient with pneumonia
50 volunteers with the symptoms of pneumonia are aged 30-70 years, and have unlimited occupation, and are diagnosed as pneumonia through imaging examination. Women are not in gestation or lactation. The staff hosts the registration, dispenses the finished product of the kit, and carries out operation explanation demonstration. The test operation was then performed by the volunteer as described in the text of the present specification, and the result was reported to the staff. And after comparison and confirmation, carrying out result statistical analysis on the staff. The results show that 50 patients can observe the color development in the T area, and the lung inflammation is prompted to be consistent with the imaging diagnosis result, and advice is kept in good compliance with the doctor's advice. Bluish black color can be observed in T areas of 28 patients, which indicates that the lung has injury bleeding and severe pneumonia symptoms, and the patients are recommended to go to a professional institution for deep examination and diagnosis. No color development was observed in the patient T-zone, and allergic/eosinophilic factors were excluded.
The invention also provides an analysis example of the use test condition of the patient with upper respiratory tract inflammation
30 patients suffering from common cold, cough, rhinitis and sphagitis are respectively collected, the ages are 20-50 years, the occupation is unlimited, the proportion of men and women is about 50%, the patients take comprehensive physical examination within one year, no other abnormal conditions, no medical history in heavy smoking or severe pollution industries, no pulmonary medical history such as lung cancer or pneumonia, and no family history of related diseases. Women are not in gestation or lactation. The staff hosts the registration, dispenses the finished product of the kit, and carries out operation explanation demonstration. After volunteers cleaned the nasal discharge, and were lightly couched to clear the throat and eliminate the expectoration, the test procedure was performed as described in the text of this specification and the results reported to the staff. And after comparison and confirmation, carrying out result statistical analysis on the staff. The results showed that no color development was observed in the T-zone of 90 total test cases of the kit, and that the risk of pneumonia was low in the tested volunteers.
The invention also provides an application example of the pneumonitis examination screening
A staff is arranged in the physical examination center and is responsible for registering forms and distributing and recycling the kit, necessary answering guidance is carried out when a subject is in question, propaganda explanation videos are rolled and played, the using method of the kit is the same as that of a specification text, and the physical examination subject can take about 5 minutes on average along with the measurement. Multiple persons can be self-tested in parallel when the persons arrive at the scene, and 30 persons can only take about 10 minutes for detection. From this estimate, detection was completed about 1440 times for 1 day (8 hours). While imaging generally takes 3-5 minutes, the subject is usually queued up, 30 people take about 150 minutes to test, and only 96 people can be completed in 8 hours. Therefore, the invention can improve the screening efficiency of physical examination by about 15 times.
The invention also provides an application example of the senile pneumonia screening
The symptoms of fever, general aching pain, headache, muscle weakness, anorexia and the like are collected in 60 cases of the aged over 60 years, the proportion of men and women is about 50%, staff host registration, and a finished product of the kit is issued and operation explanation demonstration is carried out. If the throat has foreign body sensation, the throat is lightly cough and clear, and after expectoration is eliminated, the detection operation is carried out according to the method described in the text of the specification, and the result is reported to staff. And after comparison and confirmation, carrying out result statistical analysis on the staff. The results showed that 16 of the 60 cases showed a blue-black to black-brown color in the T-zone, suggesting a risk of pneumonia, suggesting a visit to a professional for pneumonia-related examination and diagnosis. Part of the old people have no obvious fever and other symptoms when suffering from infection pneumonia, only have the symptoms of general weakness, appetite reduction and the like, are easy to ignore in early stage, and when the old people find that the lung inflammation is serious, the lung is easy to appear white, and the like. Early intervention in pneumonia is beneficial to alleviating symptoms and curing, but CT screening is limited by equipment conditions and is inconvenient for the aged to participate. The kit disclosed by the invention is convenient and easy to master in use process, and is beneficial to promoting the old people to participate in early screening and early diagnosis and early treatment of pneumonia.
The invention also provides an application example of the kit in communities
Putting 60 sets of finished products of the kit in a test point community supermarket, pasting propaganda posters with two-dimension codes, enabling a user to scan the codes by a mobile phone to watch video explanation, acquiring the kit on site to automatically detect after online registration, performing detection operation according to the method disclosed by the specification of the invention, and updating the detection result online. Summarizing and counting detection results, counting positive according to the reported results of users, and counting negative according to the reported results of users and not reported results; the logical basis is positive, so that the user's communication desire can be stimulated, the report missing rate is very low, and negative can reduce the user's communication desire, and the report missing rate is high. Statistics show that there are 5 cases of positives, the positive rate is 8.3%, and the positive rate is close to 8% of pneumonia in clinical cases of the Omikovia in 2022. The basic medical and health institutions usually lack examination and diagnosis professional facilities such as CT, and residents need to go to secondary and above hospitals for examination, which is not beneficial to pneumonia prevention and treatment. The kit is convenient to popularize and use in basic medical and health institutions, and can shunt patients with upper respiratory tract infection in communities, so that overcrowding and tension of central hospitals are avoided.
The invention also provides an application example for the preservation and stability test of the kit
The preservation condition of the kit is that the room temperature is 25+/-5 ℃ or the refrigeration temperature is low, and the humidity is less than or equal to 60%. The negative reference is prepared by mixing airway mucus samples of healthy volunteers and sub-packaging according to 1 ml/part, and the standard absorbance method measurement shows that the H2O2 content is less than 1mM, the nitrite content is less than 1mM and the Fe3+ content is less than 1mM. The positive reference is prepared by mixing airway mucus samples of volunteers of patients with pneumonia and sub-packaging according to 1 ml/part, and the standard absorbance method measurement shows that at least one index of H2O2 content, nitrite content and Fe3+ content is more than 1mM.
The repetitive references were 2mM H2O2, 2mM NaNO2, 3mM FeCl3, and the limit of detection references were 1mM H2O2, 1mM NaNO2, 1mM FeCl3. The detection method is carried out according to instructions. Each batch was tested 3 times each, 5 times at the limit of detection, and 2 batches per month. After 4 times of detection in the 2 nd month, preliminary statistics show that the detection limit coincidence rate of the kit negative/positive/repeatability reaches 100%, and meets the quality requirement.
The invention also provides test cases for portability and transport stability of the kit
Taking 3 batches of kits, storing at a high temperature of 40 ℃ for 7 days in each batch, and then, falling the kits from 1.5m height to enable the kits to fall freely, repeating the shaking and falling test for 3 times, and after observing whether the external package, the internal package and the like of the kits are damaged, testing by using a negative reference, a positive reference and a detection limit reference. Results: the kit has no damage to the outer package part and the inner package, and detection statistics show that the detection limit coincidence rate of the female/male/repeatability reaches 100% and meets the quality requirement.
The foregoing is merely illustrative of specific embodiments of the invention, and the scope of the invention is not limited thereto, but is intended to cover any variations or alternatives not contemplated by the inventors. Therefore, the protection scope of the invention should be subject to the protection scope defined by the claims.
Claims (9)
1. The utility model provides a joint detection kit of iron ion, active oxygen and nitrite, includes the test paper strip, its characterized in that: the test paper strip is provided with a detection indicator in a drying way, wherein the detection indicator comprises 2.0% -8.0% (w/v) of starch, 100-800mM of potassium ferrocyanide, 1.0-7.0M of potassium iodide, 50-300mM of calcium chloride, 0.5-3.0mM of pH regulator, 1-20mM of sodium thiosulfate and 0.5% -2.0% (w/v) of polyoxyethylene lauryl ether.
2. The kit for the combined detection of iron ions, active oxygen and nitrite according to claim 1, wherein: the device comprises a detection plate, a sampling tube, a sample extraction liquid, a test strip, a quality control indicator and a middle line reagent, wherein the sample extraction liquid is arranged in the sampling tube, the test strip is arranged in the detection plate, and the test strip is also provided with the quality control indicator and the middle line reagent in a drying way.
3. The combined detection kit for iron ions, active oxygen and nitrite according to claim 2, wherein: the composition of the sample extract comprises 50-500mM oxalic acid, 50-300mM sodium chloride and 0.5-2mM 8-hydroxyquinoline.
4. The combined detection kit for iron ions, active oxygen and nitrite according to claim 2, wherein: the alkaline pH regulator comprises one or more of sodium acetate, sodium bicarbonate, tris (hydroxymethyl) aminomethane, sodium carbonate and sodium hydroxide.
5. The combined detection kit for iron ions, active oxygen and nitrite according to claim 2, wherein: the composition of the quality control indicator comprises 5-30mM neutral red and 10-60mM alkaline pH regulator.
6. The combined detection kit for iron ions, active oxygen and nitrite according to claim 2, wherein: the composition of the intermediate line reagent comprises 5% -15% (w/v) polyoxyethylene lauryl ether and 5% -15% (w/v) sodium dodecyl sulfate.
7. The combined detection kit for iron ions, active oxygen and nitrite according to claim 2, wherein: the concentrations of the detected iron ions, active oxygen and nitrite ions are not less than 1mM, respectively.
8. The combined detection kit for iron ions, active oxygen and nitrite according to claim 2, wherein: the using method of the kit comprises the following steps:
s1: sampling, namely sampling the oral cavity by using a sampler;
s2: adding a sample, immersing the sampling end of the sampled sampler into a sample extraction liquid in a sampling tube, and mixing;
s3, pipetting, namely transferring the mixed sample extract to a test strip of a detection plate;
s4, performing S4; and (3) observing, standing until the quality control indicator on the test strip changes color, observing the color of the detection indicator on the test strip, and judging the detection result according to the color.
9. The kit for combined detection of iron ions, active oxygen and nitrite according to claim 8, wherein: the quality control indicator in the step S4 changes color to purple, and the detection indicator shows that blue-black to black-brown is a positive result, so that the contents of iron ions, active oxygen and nitrite reach or exceed a detection threshold value; and if the detection indicator does not develop color or the color is light and is a negative result, the content of iron ions, active oxygen and nitrite is lower than the detection threshold value.
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CN202311624350.0A CN117647641A (en) | 2023-11-30 | 2023-11-30 | Combined detection kit for iron ions, active oxygen and nitrite |
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