CN111808921A - Trinder reaction-based detection kit and application thereof - Google Patents

Trinder reaction-based detection kit and application thereof Download PDF

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Publication number
CN111808921A
CN111808921A CN202010542576.6A CN202010542576A CN111808921A CN 111808921 A CN111808921 A CN 111808921A CN 202010542576 A CN202010542576 A CN 202010542576A CN 111808921 A CN111808921 A CN 111808921A
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reagent
detection kit
peroxidase
kit
trinder reaction
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梁艳
赵畅
龚婷
吴年芬
凡速朋
舒芹
张雪娇
赵愿安
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Wuhan Life Origin Biotech Joint Stock Co ltd
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Wuhan Life Origin Biotech Joint Stock Co ltd
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Abstract

The invention discloses a Trinder reaction-based detection kit and application thereof, belonging to the technical field of clinical chemistry detection and comprising an R1 reagent and an R2 reagent, wherein the R1 reagent and the R2 reagent contain enzymes for catalyzing a substance to be detected to generate hydrogen peroxide, the R1 reagent also contains a buffer solution, a preservative and peroxidase, the R2 reagent contains a buffer solution, a preservative, peroxide and a chromogen substrate, the concentration of the peroxidase in the R1 reagent is 100-150 KU/L, and the R1 reagent also contains 4-aminoantipyrine with the concentration of 0.5-1 g/L. Wherein, the peroxidase and the 4-aminoantipyrine in the R1 reagent can remove clinical drugs which can participate in the Trinder reaction in a substance to be tested before the Trinder reaction, so that the determination result is more accurate. According to the invention, other substances are not required to be additionally added into the reaction system, and other impurities are not introduced; the method is simple and efficient, can be widely used for various clinical chemical detections, avoids the interference of clinical drugs, enhances the accuracy of detection results, and simultaneously improves the functional sensitivity of the kit.

Description

Trinder reaction-based detection kit and application thereof
Technical Field
The invention belongs to the technical field of clinical chemical detection, and particularly relates to a Trinder reaction-based detection kit and application thereof.
Background
The Trinder reaction is also called as coupled end point colorimetry, and the principle is that hydrogen peroxide (H) is generated by a substance to be detected through the action of enzyme2O2) In the presence of 4-aminoantipyrine (4-AAP) and Peroxidase (POD), red quinonimine compound can be generated, and the concentration of the substance to be detected can be calculated by measuring the absorbance difference before and after the reaction. The Trinder reaction is applied to many clinical biochemical detection projects, such as glucose, triglyceride, cholesterol, uric acid, high-density lipoprotein, low-density lipoprotein, free fatty acid and the like. As disclosed in Chinese patent CN106191213BThe reaction principle of the kit is that free fatty acid in human serum and coenzyme A react under the action of acetyl coenzyme A synthetase to generate fatty acyl coenzyme A, and fatty acyl coenzyme A generates H under the action of acetyl coenzyme A oxidase2O2Subsequently, a colored substance was generated by the Trinder reaction under the action of Peroxidase (POD) to determine the free fatty acid content.
However, with the rapid development of medicine, more and more medicines and test items are applied to clinic, and research shows that when a patient takes 1 kind of medicine, the percentage of the experiment interfered by the medicine is 7%, when taking 2 kinds of medicines, the percentage is 16.7%, when taking three or four kinds of medicines, the percentage is 66.7%, and when taking 5 kinds of medicines, the ratio of the interfered experiment is 100%. The influence of the drug on the detection result not only causes wrong interpretation and misdiagnosis of the detection result, but also increases some unnecessary examinations, so that the problem of drug interference in the detection is very important to solve. Clinically common drugs are: calcium dobesilate (used for treating microvascular diseases and the like), etamsylate (a hemostatic drug), methyldopa (an antihypertensive drug), levodopa (used for treating parkinsonism), dopamine (a nerve conduction substance) and the like, because of strong reducibility or similarity to a chromogen substrate structure in a Trinder reaction, hydrogen peroxide is consumed in the Trinder reaction, and the measured value of an object to be measured is low. And the drugs are stable in structure and are not easy to damage or degrade, so that the interference on the clinical chemical detection based on the Trinder reaction can be generated, and the determination result is influenced.
Disclosure of Invention
Aiming at the defects in the prior art, the invention provides a Trinder reaction-based detection kit, which removes clinical drugs participating in Trinder reaction in an object to be detected by adding peroxidase and 4-aminoantipyrine in advance into an R1 reagent, thereby avoiding the interference of various clinical drugs on Trinder reaction detection in the chemical detection process.
In order to achieve the purpose, the invention adopts the technical scheme that:
a detection kit based on Trinder reaction comprises an R1 reagent and an R2 reagent, wherein the R1 reagent and the R2 reagent contain enzymes for catalyzing a substance to be detected to generate hydrogen peroxide, the R1 reagent also contains a buffer solution, a preservative and peroxidase, the R2 reagent contains a buffer solution, a preservative, peroxide and a chromogen substrate, the concentration of the peroxidase in the R1 reagent is 100-150 KU/L, and the R1 reagent also contains 4-aminoantipyrine with the concentration of 0.5-1 g/L;
the detection kit can improve the capacity of resisting interference of clinical drugs, wherein the clinical drugs comprise: one or more of calcium dobesilate, etamsylate, methyldopa, levodopa and dopamine.
The technical scheme of the invention also provides a method for improving the anti-interference capability of Trinder reaction detection, which comprises the following steps:
s1, adding peroxidase to a final concentration of 100-150 KU/L before the Trinder reaction, and simultaneously adding 4-aminoantipyrine to a final concentration of 0.5-1 g/L to remove clinical drugs in the object to be detected, wherein the clinical drugs can participate in the Trinder reaction;
s2, adding an enzyme capable of catalyzing the object to be detected to generate hydrogen peroxide during the Trinder reaction, and simultaneously adding peroxidase and aniline derivatives to the final concentrations of 25-30 KU/L and 1g/L respectively to perform the Trinder reaction to generate a color substance;
and S3, measuring the absorbance before and after the Trinder reaction to detect the concentration of the substance to be detected.
The technical scheme of the invention also provides the application of the Trinder reaction-based detection kit in clinical chemical detection.
Compared with the prior art, the invention has the beneficial effects that: peroxidase and 4-aminoantipyrine are added into an R1 reagent before Trinder reaction to consume clinical drugs in a substance to be detected, so that the interference of the clinical drugs on the Trinder reaction detection is avoided, the determination result is more accurate, the excessive peroxidase and 4-aminoantipyrine can be directly used for subsequent Trinder reaction, and other substances are not required to be additionally added into a reaction system in the invention, so that other impurities are avoided from being introduced. The method is simple and efficient, can be widely used for various clinical chemical detections and preparation of corresponding clinical chemical detection kits, enhances the accuracy of detection results, and avoids drug interference.
Detailed Description
The invention provides a Trinder reaction-based detection kit, which consists of an R1 reagent and an R2 reagent, wherein the R1 reagent and the R2 reagent contain enzymes for catalyzing a substance to be detected to generate hydrogen peroxide, the R1 reagent also contains a buffer solution, a preservative and peroxidase, the R2 reagent contains a buffer solution, a preservative, peroxide and a chromogen substrate, the concentration of the peroxidase in the R1 reagent is 100-150 KU/L, and the R1 reagent also contains 4-aminoantipyrine with the concentration of 0.5-1 g/L;
the detection kit can improve the anti-interference capability of clinical medicines, and the clinical medicines comprise: one or more of calcium dobesilate, etamsylate, methyldopa, levodopa and dopamine.
The catalytic enzymes used in the detection kit are different for different analytes, and the catalytic reaction can be started only after the reagents R1 and R2 are mixed, so that the analytes are gradually catalyzed to generate hydrogen peroxide.
According to the detection kit, peroxidase and 4-aminoantipyrine are added into an R1 reagent to remove clinical drugs participating in Trinder reaction in a substance to be detected. Part of clinical drugs have strong reducibility or are similar to chromogen substrate structures in Trinder reaction, so that the clinical drugs can participate in the Trinder reaction and compete to consume hydrogen peroxide generated by a substance to be detected, and the measured value of the final substance to be detected is low. In order to avoid the influence of clinical drugs on the determination result, the invention adds excessive peroxidase and 4-aminoantipyrine into the R1 reagent to process a serum sample before the Trinder reaction, so that the clinical drugs react with the hydrogen peroxide in the serum, the peroxidase in the R1 reagent and the 4-aminoantipyrine, and the clinical drugs are consumed in advance, thereby ensuring more accurate determination result and avoiding the interference of the clinical drugs on the Trinder reaction detection.
Preferably, the buffer solution in the R1 reagent and the R2 reagent is any one of phosphate buffer solution, HEPES buffer solution, Good's buffer solution, MOPSP buffer solution, BICINE buffer solution and glycine buffer solution.
Preferably, the concentration of the buffer is 20-100 mmol/L.
Preferably, the preservative in the R1 reagent and the R2 reagent is one or more of sodium azide, dichloroacetamide, imidazolidinyl urea, thimerosal, gentamicin, isothiazolinone, potassium sorbate, sodium benzoate, paraben, Proclin series and KroVin series.
Preferably, the concentration of the preservative is 0.1-10 g/L.
Preferably, the R1 reagent also comprises a metal ion chelating agent and a surfactant for maintaining the stability of the peroxidase.
Preferably, in the R2 reagent, the concentration of the peroxidase is 25-30 KU/L.
Preferably, the chromogen substrate is an aniline derivative, the concentration of the aniline derivative is 1g/L, and the aniline derivative is used as the chromogen substrate, so that compared with the traditional phenol substance, the stability and the solubility of a chromogen group, the sensitivity of a product and the stability of color and luster are improved.
Preferably, the aniline derivative is one or more of DHBS, TOOS, HDAOS, F-DAOS, DHBA, TOPS, ADOS, ADPS, ALPS, DAOS, MADB, MAOS and TODB. Wherein DHBS is sodium 3, 5-dichloro-2-hydroxybenzenesulfonate, TOOS is N-ethyl-N- (2-hydroxy-3-sulfopropyl) -3-methylaniline sodium salt, HDAOS is N- (2-hydroxy-3-sulfopropyl) -3' 5-dimethoxyaniline sodium salt, F-DAOS is N-ethyl-N- (2-hydroxy-3-sulfopropyl) -3, 5-dimethoxy-4-fluoroaniline, DHBA is 3, 4-dihydroxybenzylamine hydrobromide, TOPS is N-ethyl-N- (3-sulfopropyl) -3-toluidine sodium salt, ADOS is N-ethyl-N- (2-hydroxy-3-sulfopropyl) -3-methoxyaniline sodium salt, ADPS is N-ethyl-N- (3-sulfopropyl) -3-methoxyaniline sodium salt, ALPS is N-ethyl-N- (3-sulfopropyl) aniline sodium salt, DAOS is N-ethyl-N- (2-hydroxy-3-sulfopropyl) -3, 5-dimethoxyaniline sodium salt, MADB is N, N-bis (4-sulfopropyl) -3, 5-dimethylaniline sodium salt, MAOS is N-ethyl-N- (2-hydroxy-3-sulfopropyl) -3, 5-dimethylaniline sodium salt, TODB is N, N-bis (4-sulfobutyl) -3-methylaniline disodium salt, and DHBS has a better color development effect.
Preferably, the kit comprises: the kit comprises a creatinine detection kit (enzyme method), a triglyceride detection kit (GPO-PAP method), a total cholesterol detection kit (COD-PAP method), a free fatty acid detection kit (enzyme method), a glycated albumin detection kit (enzyme method), a high-density lipoprotein cholesterol detection kit, a low-density lipoprotein cholesterol detection kit, a uric acid detection kit (enzyme method), a glucose detection kit (GPO-PAP method), an adenosine deaminase detection kit, a lipase detection kit, a 5' -nucleotidase detection kit and the like.
The invention also provides a method for improving the anti-interference capability of Trinder reaction detection by using the detection kit based on the Trinder reaction, which comprises the following steps:
s1, adding peroxidase to a final concentration of 100-150 KU/L before the Trinder reaction, and simultaneously adding 4-aminoantipyrine to a final concentration of 0.5-1 g/L to remove clinical drugs in the object to be detected, wherein the clinical drugs can participate in the Trinder reaction;
s2, adding an enzyme capable of catalyzing the object to be detected to generate hydrogen peroxide during the Trinder reaction, and simultaneously adding peroxidase and aniline derivatives to the final concentrations of 25-30 KU/L and 1g/L respectively to perform the Trinder reaction to generate a color substance;
and S3, measuring the absorbance before and after the Trinder reaction to detect the concentration of the substance to be detected.
The technical solutions of the present invention will be described clearly and completely with reference to the following embodiments of the present invention, and it should be understood that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The performance evaluation method for each kit in the embodiment of the invention is concretely as follows:
(1) precision: repeatedly measuring the same sample for 10 times by adopting each kit, calculating the coefficient of variation (CV,%) and recording the coefficient of variation as the precision, wherein the precision is less than or equal to 5 percent and is regarded as meeting the requirement;
(2) linear range: determining samples in a certain concentration range by using each kit, and calculating a linear correlation coefficient r, wherein r is more than or equal to 0.99 and is regarded as meeting the requirement;
(3) accuracy: measuring a third-party quality control product by using each kit, and calculating the deviation between the measured value of each kit and the target value of the third-party quality control product, wherein the deviation is less than or equal to 10 percent to meet the requirement;
(4) functional sensitivity: adopting each kit to measure a low-concentration sample, measuring and calculating the diurnal variation coefficient of each kit, wherein the average concentration of the corresponding detection sample when the diurnal variation coefficient is 20 percent is recorded as the functional sensitivity;
(5) stability: each kit was left at 37 ℃ for 7 days, and the above-mentioned indexes were evaluated: precision, linear range, accuracy, functional sensitivity;
(6) interference evaluation: the interfering substances were diluted with ultrapure water in a gradient, and the concentrations of the interfering substances were added to the blank control serum (i.e., normal human serum mixture containing no interfering substances) at a ratio of 1:9, respectively, to give final concentrations of the interfering substances as shown in the table in the examples. The absorbance values of the blank serum and the serum added with the interfering substances with different concentrations are respectively measured by using each kit, each serum sample is measured for 3 times to obtain a mean value, and the deviation between the mean value and the measured value of the blank control serum ((mean value-blank control)/blank control x 100%) is calculated, namely the interference degree. If the absolute value of the interference degree is less than 10%, recording as anti-interference, otherwise, recording as non-anti-interference. Wherein the interfering drugs comprise calcium dobesilate, etamsylate, methyldopa, levodopa, and dopamine; common interfering substances in serum are ascorbic acid, bilirubin, hemoglobin and triglycerides.
Example 1 Creatinine detection kit
The embodiment provides a creatinine detection kit, which is compared with a conventional creatinine detection kit, wherein the detection kit is a test group I, the conventional creatinine detection kit is a control group I, and the formula is as follows:
Figure BDA0002539505360000061
compared with a control group I, 150KU/L of peroxidase and 1g/L of 4-aminoantipyrine are added into an R1 reagent component for consuming the drug, and simultaneously, because the stability of the peroxidase is lower than that of catalase, the test group I also adds a proper amount of metal ion chelating agent HEDTA and surfactant GENAP-080 into an R1 reagent component for ensuring the stability of the peroxidase; the first test group also added 1g/L DHBS to the R2 reagent component instead of TOOS in the first R1 reagent component as a chromogen substrate.
Taking blank sample (ultrapure water), sample to be tested and creatinine calibrator (from Wuhan's Source science and technology Co., Ltd.) each 6 μ L, adding 270 μ L of R1 reagent, mixing, keeping at 37 deg.C for 5min, adjusting to zero with the blank sample, measuring absorbance of the sample to be tested at 546nm wavelength by using HITACHI 7180 (from Wuhan's Source science and technology Co., Ltd.), and recording as A1Then, 90. mu.L of R2 reagent was added to each tube, mixed well, and incubated at 37 ℃ for 5min, zeroed with a blank sample, calibrated with creatinine calibrator, and measured for absorbance at 546nm of the sample to be tested using HITACHI 7180 (available from Softscience, Inc., Wuhan, Ltd.) as A2That is, the absorbance Δ A of the sample to be measured is equal to A2-A1. The properties of the control group I and the test group I were measured in accordance with the above-mentioned property evaluation methods (linear range portion, concentration range of the sample: 2.4. mu. mol/L-8840.0. mu. mol/L), and the results are shown in tables 1 to 3.
TABLE 1 drug interference bias data
Figure BDA0002539505360000071
Figure BDA0002539505360000081
Figure BDA0002539505360000091
TABLE 2 interference bias data for general interfering substances in serum
Figure BDA0002539505360000092
Figure BDA0002539505360000101
Table 3 performance evaluation data
Figure BDA0002539505360000102
Figure BDA0002539505360000111
According to the measurement results in tables 1-2, the creatinine detection kits obtained from the control group one and the test group one have anti-interference capability (the absolute value of interference degree is less than or equal to 10%) for common interference substances (bilirubin, hemoglobin, ascorbic acid and triglyceride) in serum, but the creatinine detection kits obtained from the control group one have no anti-interference capability for common clinical drugs (calcium dobesilate, etamsylate, methyldopa, levodopa and dopamine), and the creatinine detection kits prepared by the method of the present invention have strong anti-interference capability (the interference degree is less than 5%) for common clinical drugs. According to the determination results in table 3, after the reagent prepared by the first test group is placed at 37 ℃ for 7 days, the precision, the linear correlation coefficient and the accuracy of the reagent can meet the requirements, and the reagent does not change greatly, the change difference is within an acceptable range, namely, the stability is stronger, and the functional sensitivity of the kit prepared by the invention is obviously higher than that of the first control group, namely, when the concentration of the object to be detected is lower, the first test group can also accurately measure the concentration of the object to be detected, and the sensitivity is obviously enhanced. Namely, the creatinine detection kit which has strong anti-interference capability to common clinical drugs, improved functional sensitivity and no great change in other properties is prepared in the embodiment.
Example 2 detection kit for free fatty acids
The embodiment provides a free fatty acid detection kit, and the kit is compared with a conventional free fatty acid detection kit, wherein the detection kit is a second test group, the conventional creatinine detection kit is a second control group, and the formula is as follows:
Figure BDA0002539505360000112
Figure BDA0002539505360000121
compared with a control group II, the test group II is added with 100KU/L of peroxidase and 0.5g/L of 4-aminoantipyrine for consuming drugs in the R1 reagent component, and the R1 reagent does not contain a metal ion chelating agent and a surfactant; 1g/L of HAOS was added to the R2 reagent component in place of TOOS in the control-R1 reagent component as a chromogen substrate. The assay was performed as in example 1, with each sample size being 5. mu.L, and 240. mu.L of R1 reagent and 60. mu.L of R2 reagent were added. The properties of the control group II and the test group II were measured by the aforementioned property evaluation methods (linear range portion, concentration range of the sample: 0.010mmol/L-3.000mmol/L), and the results are shown in tables 4-6.
Table 4 drug interference bias data
Figure BDA0002539505360000122
Figure BDA0002539505360000131
Figure BDA0002539505360000141
TABLE 5 interference bias data for general interfering substances in serum
Figure BDA0002539505360000142
Figure BDA0002539505360000151
Figure BDA0002539505360000161
Table 6 performance evaluation data
Figure BDA0002539505360000162
According to the measurement results in tables 4 to 5, the free fatty acid detection kits obtained from the second control group and the second test group have anti-interference capability (the absolute value of interference degree is less than or equal to 10%) on common interference substances (bilirubin, hemoglobin, ascorbic acid and triglyceride) in serum, but the free fatty acid detection kits of the second control group have no anti-interference capability on common clinical medicines (calcium dobesilate, etamsylate, methyldopa, levodopa and dopamine), and the free fatty acid detection kits prepared by the invention have strong anti-interference capability (the interference degree is less than 5%) on common clinical medicines.
From the measurement results in table 6, it can be seen that, since the R1 reagent of the second test group does not contain a metal ion chelating agent and a surfactant, the difference in the precision, linear correlation coefficient and accuracy of the prepared reagent before and after being placed at 37 ℃ for 7 days is slightly larger than that of the reagent prepared in the first test group of example 1, but still satisfies the requirement that the R1 reagent can still maintain strong stability even if the R1 reagent does not contain a metal ion chelating agent and a surfactant, and the functional sensitivity of the reagent prepared by the present invention is significantly higher than that of the second control group. Namely, the free fatty acid detection kit which has strong anti-interference capability to common clinical drugs, is improved in functional sensitivity and has no great change in other performances is prepared in the embodiment.
Example 3 Triglycerides detection kit
The present embodiment provides a triglyceride detection kit, and compares the triglyceride detection kit with a conventional triglyceride detection kit, wherein a test group three is the detection kit of the present invention, a control group three is the conventional triglyceride detection kit, and a formula is specifically shown as follows:
Figure BDA0002539505360000171
compared with a third control group, the third test group adds 4mmol/L (0.81g/L) of 4-aminoantipyrine into the R1 reagent component, namely, peroxidase and 4-aminoantipyrine are added simultaneously before the Trinder reaction, the third control group adds 4-aminoantipyrine into the R2 reagent component for carrying out the Trinder reaction, and the third test group adds DHBS into the R2 reagent component to replace parachlorophenol in the third R1 reagent component of the control group as a chromogen substrate. The assay was performed as described in example 1, wherein the amount of each sample was 3. mu.L, and 240. mu.L of R1 reagent and 60. mu.L of R2 reagent were added. The properties of the control group III and the test group III were measured in accordance with the above-mentioned property evaluation methods (linear range portion, concentration range of the sample: 0.05mmol/L to 10.00mmol/L) and the results are shown in tables 7 to 9.
TABLE 7 drug interference bias data
Figure BDA0002539505360000181
Figure BDA0002539505360000191
TABLE 8 interference bias data for general interfering substances in serum
Figure BDA0002539505360000201
Figure BDA0002539505360000211
TABLE 9 evaluation data of Performance
Figure BDA0002539505360000212
According to the determination results in tables 7-8, the triglyceride detection kits obtained from the third control group and the third test group have anti-interference capability (the absolute value of interference degree is less than or equal to 10%) on common interfering substances (bilirubin, hemoglobin and ascorbic acid) in serum, but the triglyceride detection kits obtained from the third control group have no anti-interference capability on common clinical drugs (calcium dobesilate, etamsylate, methyldopa, levodopa and dopamine), while the triglyceride detection kits prepared according to the invention have strong anti-interference capability (the interference degree is less than 5%) on common clinical drugs, namely, when peroxidase and 4-aminoantipyrine exist in the R1 reagent simultaneously (when peroxidase and 4-aminoantipyrine are added before Trinder reaction), drug consumption can be achieved, and the effect of drug interference on Trinder reaction detection can be avoided, the effect of drug consumption cannot be achieved by only adding peroxidase. According to the determination results in table 9, after the reagent prepared by the third test group is placed at 37 ℃ for 7 days, the precision, the linear correlation coefficient and the accuracy of the reagent can meet the requirements, and the reagent has no large change, the change difference is within an acceptable range, namely, the stability is stronger, and the functional sensitivity of the reagent prepared by the invention is obviously higher than that of the third control group. Namely, the triglyceride detection kit which has strong anti-interference capability to common clinical drugs, improved functional sensitivity and no great change in other properties is prepared in the embodiment.
The peroxidase and 4-aminoantipyrine in the R1 reagent can react with a plurality of common clinical drugs which have strong reducibility in serum to be detected or have similar structures with chromogen substrates in Trinder reaction so as to improve the anti-interference capability of the kit, so the method can be widely used for preparing a plurality of detection kits based on the Trinder reaction.
Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.

Claims (10)

1. A Trinder reaction-based detection kit comprises an R1 reagent and an R2 reagent, wherein the R1 reagent and the R2 reagent contain enzymes for catalyzing a substance to be detected to generate hydrogen peroxide, the R1 reagent also contains a buffer solution, a preservative and peroxidase, and the R2 reagent contains a buffer solution, a preservative, peroxide and a chromogen substrate, and is characterized in that the concentration of the peroxidase in the R1 reagent is 100-150 KU/L, and the R1 reagent also contains 4-aminoantipyrine with the concentration of 0.5-1 g/L;
the detection kit can improve the capacity of resisting interference of clinical drugs, wherein the clinical drugs comprise: one or more of calcium dobesilate, etamsylate, methyldopa, levodopa and dopamine.
2. The Trinder reaction-based assay kit according to claim 1, wherein the R1 reagent further comprises a metal ion chelating agent and a surfactant.
3. The Trinder reaction-based detection kit according to claim 1, wherein the concentration of peroxidase in the R2 reagent is 25-30 KU/L.
4. The Trinder reaction-based assay kit according to claim 1, wherein the chromogen substrate is an aniline derivative, and the concentration of the aniline derivative is 1 g/L.
5. The Trinder reaction-based assay kit according to claim 4, wherein the aniline derivative is one or more of DHBS, TOOS, HDAOS, F-DAOS, DHBA, TOPS, ADOS, ADPS, ALPS, DAOS, MADB, MAOS, and TODB.
6. The Trinder reaction-based assay kit according to claim 1, wherein the kit comprises: the kit comprises a creatinine detection kit, a triglyceride detection kit, a total cholesterol detection kit, a free fatty acid detection kit, a glycated albumin detection kit, a high density lipoprotein cholesterol detection kit, a low density lipoprotein cholesterol detection kit, a uric acid detection kit, a glucose detection kit, an adenosine deaminase detection kit, a lipase detection kit and a 5' -nucleotidase detection kit.
7. A method for improving the anti-interference capability of Trinder reaction detection is characterized by comprising the following steps:
s1, adding peroxidase to a final concentration of 100-150 KU/L before the Trinder reaction, and simultaneously adding 4-aminoantipyrine to a final concentration of 0.5-1 g/L to remove clinical drugs in the object to be detected, wherein the clinical drugs can participate in the Trinder reaction;
s2, adding an enzyme capable of catalyzing the object to be detected to generate hydrogen peroxide during the Trinder reaction, and simultaneously adding peroxidase and aniline derivatives to the final concentrations of 25-30 KU/L and 1g/L respectively to perform the Trinder reaction to generate a color substance;
and S3, measuring the absorbance before and after the Trinder reaction to detect the concentration of the substance to be detected.
8. A creatinine assay kit, wherein said creatinine kit comprises R1 reagent and R2 reagent, wherein said R1 reagent comprises: 100mmol/L of Good's buffer solution, 20KU/L of sarcosine oxidase, 50KU/L of creatine amidinohydrolase, 10KU/L of ascorbic acid oxidase, 150KU/L of peroxidase, 1g/L of 4-aminoantipyrine, 0.5g/L, Proclin3001g/L, GENAPOLX-08010g/L of HEDTA, and pH is 7.5; the R2 reagent includes: good's buffer 50mmol/L, creatinine amidohydrolase 300KU/L, potassium ferrocyanide 0.05g/L, sodium azide 1g/L, peroxidase 30KU/L, DHBS1g/L, pH 7.5.
9. A free fatty acid detection kit, which is characterized by consisting of an R1 reagent and an R2 reagent, wherein the R1 reagent comprises: 50mmol/L phosphate buffer solution, 100KU/L peroxidase, 4g/L coenzyme A, 6g/L, Proclin3001g/L adenosine triphosphate, 0.5 g/L4-aminoantipyrine, and pH of 7.0; the R2 reagent includes: 50mmol/L phosphate buffer solution, 50KU/L fatty acyl-CoA oxidase, 5KU/L fatty acyl-CoA synthetase, 25KU/L peroxidase, 1g/L, HDAOS1 sodium azide, 1g/L, and pH 7.0.
10. A triglyceride test kit, which is characterized by consisting of an R1 reagent and an R2 reagent, wherein the R1 reagent comprises: HEPES buffer solution 50mmol/L, glycerol kinase 18KU/L, peroxidase 130KU/L, glycerol phosphate oxidase 18KU/L, sodium cholate 5mmol/L, PC-3001g/L, 4-aminoantipyrine 4mmol/L, and pH 6.5; the R2 reagent includes: HEPES buffer solution 50mmol/L, lipoprotein lipase 18KU/L, sodium azide 1g/L, adenosine triphosphate 0.15mmol/L, oxamic acid 0.05mmol/L, peroxidase 30KU/L, DHBS1g/L, pH 6.5.
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Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112608976A (en) * 2020-12-18 2021-04-06 珠海市德灏生物科技有限公司 Detection kit and detection method for periodontal pathogen infection marker
CN112630216A (en) * 2020-11-27 2021-04-09 上海绅道生物科技有限公司 Kit and method for detecting 1,5-AG
CN114441516A (en) * 2021-12-20 2022-05-06 苏州百源基因技术有限公司 Uric acid detection kit and preparation method thereof
CN115201134A (en) * 2022-09-15 2022-10-18 吉林大学第一医院 Creatinine detection kit resistant to piceatannol interference and application thereof
WO2023190087A1 (en) * 2022-03-31 2023-10-05 東洋紡株式会社 Enhacement of accuracy when measuring high-creatinine-value specimen

Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4517287A (en) * 1979-04-04 1985-05-14 Boehringer Mannheim Gmbh Method and reagent for the enzymatic determination of enzyme substrates
CN102023158A (en) * 2010-10-19 2011-04-20 李立和 Method for enzymatically determining creatinine in serum by two steps
CN103913581A (en) * 2013-11-20 2014-07-09 天津市宝坻区人民医院 Cyclophorase determination method for triglyceride in serum
CN104678107A (en) * 2015-02-10 2015-06-03 浙江凯成生物科技有限公司 Method, reagent and kit for quantitatively detecting free fatty acid
JP2015204825A (en) * 2014-04-10 2015-11-19 ヤマサ醤油株式会社 L-glutamine measurement kit
CN106290323A (en) * 2015-06-04 2017-01-04 章丘美高义医疗器械有限公司 A kind of stable, uric acid reagent that capacity of resisting disturbance is strong and detection method
EP3258273A1 (en) * 2016-06-15 2017-12-20 Peking Union Medical College Hospital, Chinese Academy of Medical Sciences A kit and method for detecting creatinine

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4517287A (en) * 1979-04-04 1985-05-14 Boehringer Mannheim Gmbh Method and reagent for the enzymatic determination of enzyme substrates
CN102023158A (en) * 2010-10-19 2011-04-20 李立和 Method for enzymatically determining creatinine in serum by two steps
CN103913581A (en) * 2013-11-20 2014-07-09 天津市宝坻区人民医院 Cyclophorase determination method for triglyceride in serum
JP2015204825A (en) * 2014-04-10 2015-11-19 ヤマサ醤油株式会社 L-glutamine measurement kit
CN104678107A (en) * 2015-02-10 2015-06-03 浙江凯成生物科技有限公司 Method, reagent and kit for quantitatively detecting free fatty acid
CN106290323A (en) * 2015-06-04 2017-01-04 章丘美高义医疗器械有限公司 A kind of stable, uric acid reagent that capacity of resisting disturbance is strong and detection method
EP3258273A1 (en) * 2016-06-15 2017-12-20 Peking Union Medical College Hospital, Chinese Academy of Medical Sciences A kit and method for detecting creatinine

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112630216A (en) * 2020-11-27 2021-04-09 上海绅道生物科技有限公司 Kit and method for detecting 1,5-AG
CN112608976A (en) * 2020-12-18 2021-04-06 珠海市德灏生物科技有限公司 Detection kit and detection method for periodontal pathogen infection marker
CN114441516A (en) * 2021-12-20 2022-05-06 苏州百源基因技术有限公司 Uric acid detection kit and preparation method thereof
WO2023190087A1 (en) * 2022-03-31 2023-10-05 東洋紡株式会社 Enhacement of accuracy when measuring high-creatinine-value specimen
CN115201134A (en) * 2022-09-15 2022-10-18 吉林大学第一医院 Creatinine detection kit resistant to piceatannol interference and application thereof
CN115201134B (en) * 2022-09-15 2022-12-27 吉林大学第一医院 Creatinine detection kit resistant to piceatannol interference and application thereof

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