CN110763837A - Electronic quality control card for time-resolved fluorescence immunoassay analyzer and electronic quality control card assembly - Google Patents
Electronic quality control card for time-resolved fluorescence immunoassay analyzer and electronic quality control card assembly Download PDFInfo
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Abstract
The invention discloses an electronic quality control card for a time-resolved fluorescence immunoassay analyzer, which is obtained by breakthroughly arranging two fluorescence quality control lines on an NC membrane of the same quality control card and processing the two fluorescence quality control lines by specific sealing liquid. The obtained electronic quality control card can realize the simultaneous verification of the low value detection and the high value detection of the time-resolved fluorescence immunoassay analyzer through one-step quality control operation, eliminates the errors caused by the conventional two-time verification operation, and simplifies the verification process. The electronic quality control card has high checking precision and long service life, is not easy to be influenced by external environment to cause fluorescence attenuation, and can generate obvious economic benefit and application value.
Description
Technical Field
The invention relates to the technical field of quality control cards, in particular to an electronic quality control card for a time-resolved fluorescence immunoassay analyzer and an electronic quality control card assembly.
Background
The time-resolved fluorescence analyzer is an apparatus for quantitatively analyzing various immunodetection items in human blood and other body fluids by using a time-resolved fluorescence analysis method (TRFIA), is suitable for various inspection medicine such as infectious disease inspection, endocrinology inspection, cytology inspection, oncology inspection and the like, and has an increasingly large application space. The use of time-resolved fluoroimmunoassay analyzers requires periodic quality control, checking of the function of the optical system and the analytical processing system to ensure that the performance of the instrument is within an acceptable range, and verifying the validity of the operating results of the entire system. At present, time-resolved fluorescent microspheres are diluted to a certain concentration and then are marked on an NC membrane, and then the NC membrane is assembled in a shell of a reagent plate and is used as an electronic quality control card of a time-resolved fluorescence immunoassay analyzer. In addition, in the process of checking the analyzer, low-value quality control and high-value quality control are generally required to be performed in sequence, and certain quality control errors exist.
Disclosure of Invention
In order to solve the defects in the prior art, the invention provides the electronic quality control card for the time-resolved fluorescence immunoassay analyzer, which simultaneously carries two low-value and high-value fluorescent quality control lines on an NC membrane, realizes one-time operation and simultaneously completes low-value quality control and high-value quality control, and achieves more accurate quality control. Meanwhile, the quality control card has higher stability, the fluorescence intensity is not easy to be attenuated by the influence of the environment, and the stable and accurate test result can be kept for a long time to ensure the quality control.
In order to achieve the purpose, the electronic quality control card for the time-resolved fluoroimmunoassay analyzer comprises a quality control NC film, wherein two fluorescence quality control lines are carried on the quality control NC film;
the quality control NC membrane is prepared by the following steps:
a. obtaining a fluorescent substance line control solution;
diluting the fluorescent microspheres step by using a diluent, and performing microscopic examination after dilution to ensure that the fluorescent microspheres are not crosslinked and are in a single state to obtain fluorescent microsphere solutions with high fluorescence intensity and low fluorescence intensity respectively, namely two quality control line solutions;
the diluent is phosphate buffer solution containing 0.01-0.05% of sodium dodecyl sulfate by mass, wherein the mass concentration of substances in the phosphate buffer solution is 45 mmol/L; the fluorescent microspheres are hydrophilic time-resolved fluorescent microspheres;
b. marking a fluorescence quality control line on the NC film;
placing the quality control line solution obtained in the step a on a film scribing instrument, scribing or spraying two quality control line solutions on different positions of the surface of the NC film by the film scribing instrument to obtain a fluorescence high-value quality control line and a fluorescence low-value quality control line required by a time-resolved fluorescence immunoassay analyzer, and then drying the NC film in a drying tower;
c. sealing and drying the membrane;
c, sealing the NC membrane dried in the step b by using a sealing liquid, and drying the sealed NC membrane again to obtain a quality control NC membrane;
the sealing liquid mainly comprises a sealing component, a surfactant component and a phosphate buffer solution (PB solution) with the pH value of 7.0-8.0 and 50-60 mmol/L, wherein the sealing component is polyvinyl alcohol 9000(PVA9000), the mass percentage of the sealing component in the sealing liquid is 1.0-1.3%, the surfactant component is at least one of Pluronic F68, RHOSUREF ON-870 and TRITON X-305, and the mass percentage of the surfactant component in the sealing liquid is 0.5-1.0%.
According to the electronic quality control card, two fluorescence quality control lines, namely a fluorescence low-value quality control line and a fluorescence high-value quality control line, are arranged on the NC membrane in a breakthrough manner, so that the low value detection and the high value detection of the analyzer are simultaneously verified by one quality control card, errors caused by two verification operations of low-value quality control and high-value quality control which are respectively carried out by two quality control cards in conventional operation are eliminated, the verification process is simplified, the quality control accuracy of a time-resolved fluorescence immunoassay analyzer can be improved, and the electronic quality control card has a remarkable optimization significance. In order to avoid the problem that two fluorescence quality control lines possibly have mutual influence on the same quality control card NC membrane, the invention carries out adaptive overall improvement on the aspects of the preparation method of the quality control line solution, the membrane scribing and sealing treatment of the fluorescence quality control lines, and the like, and on the basis of the correlation mechanism based on all factors, on one hand, the preparation method of the quality control line solution is adjusted, so that the low-value intensity and the high-value intensity scribed on the same NC membrane do not generate fluorescence detection interference with each other; on the other hand, the composition of the conventional sealing liquid is changed, the isolation protection effect on the fluorescent quality control line can be exerted, the influence of the external environment on the fluorescent intensity of the quality control line is resisted, the sensitivity and attenuation to humidity, temperature and illumination are reduced, the service life of the electronic quality control card is obviously prolonged, and the verification accuracy is ensured.
As a limitation to the above technical solution, the electronic quality control card further includes a reagent case for assembling the strip-shaped quality control NC film, and a two-dimensional code pattern including quality control standard information is sprayed on the reagent case.
The marked and sealed NC film is cut into strips and assembled on a special reagent shell of a conventional electronic quality control card to form the quality control card, and a two-dimensional code pattern is attached to the reagent shell to facilitate the standard comparison of quality control data, so that the quality control verification is facilitated.
As a limitation to the above technical scheme, the hydrophilic time-resolved fluorescent microsphere is any one of a carboxyl-modified time-resolved fluorescent microsphere, a hydroxyl-modified time-resolved fluorescent microsphere, a coupled protein A time-resolved fluorescent microsphere and a coupled protein G time-resolved fluorescent microsphere.
The types of the fluorescent microspheres are further limited, and the use performance of the electronic quality control card is optimized while the preparation of the electronic quality control card is perfected.
As a limitation to the technical scheme, the scribing or spraying operation of the fluorescent quality control line on the NC film and the subsequent drying process in the step b are operated in a continuous state;
the step of continuously operating comprises:
placing the NC film roll on a take-out reel, then passing through a film scribing instrument, wherein one film scribing instrument can be used for placing a plurality of film scribing spray heads, and simultaneously scribing or spraying two quality control line solutions diluted to required concentrations on the NC film; and simultaneously scribing the rear end of the NC film, immediately feeding the front end of the scribed NC film into a drying tower preheated to the drying temperature in advance, and drying.
As a limitation to the technical scheme, the running time of each fluorescence quality control line on the NC membrane from the start of scribing to the completion of drying is 5-6 min, and the drying temperature is 30-35 ℃.
As a limitation to the above technical solution, the operation of the membrane sealing and the subsequent drying process in step c are operated in a continuous state;
the step of continuously operating comprises:
and c, enabling the NC membrane dried in the step b to pass through a soaking liquid tank with sealing liquid at a constant speed to seal the NC membrane, sealing the rear end of the NC membrane, simultaneously enabling the front end of the sealed NC membrane to immediately enter a drying tower preheated to the drying temperature in advance, and rolling after drying.
As a limitation on the technical scheme, the running time of the NC membrane from the beginning of sealing to the completion of drying is 5-6 min, and the drying temperature is 30-35 ℃.
The continuous process is adopted for scribing and sealing the NC film, so that the preparation operation is optimized, the preparation time is shortened, the checking precision of the electronic quality control card is further improved, and the product quality of the electronic quality control card is ensured.
Meanwhile, the invention also provides an electronic quality control card component for the time-resolved fluorescence immunoassay analyzer, which comprises the electronic quality control card and a storage box for storing the electronic quality control card; the storage box comprises a box body with a hollow interior, a sealable cover body is arranged at the top of the box body, an air hole with a one-way sealing valve is formed in the cover body, and a clamping groove for containing the electronic quality control card and a dehumidification groove for storing a drying agent are formed in the box body.
As a limitation to the technical scheme, the Coefficient of Variation (CV) value of the electronic quality control card assembly does not exceed 0.4% in one year.
In order to better store the electronic quality control card and prolong the service life and the use precision of the electronic quality control card, the invention also designs a storage box suitable for storing the quality control card, the quality control card is stored in a box body environment capable of storing pressure by utilizing the sealing property and the vacuumizing structure of the box body, and the drying agent is additionally arranged in the box body, so that the attenuation influence of the external environment on the fluorescence intensity of the quality control card is reduced, and the effectiveness of the quality control card is kept.
In summary, the electronic quality control card obtained by the technical scheme of the invention is used for checking the detection limit of the time-resolved fluorescence immunoassay analyzer, two fluorescence quality control lines are arranged on the NC film of the same quality control card in a breakthrough manner, and the simultaneous checking of the detection low value and the detection high value can be realized through one-step quality control operation, so that the error caused by two times of checking operations of two quality control cards in the conventional operation is eliminated, the checking process is simplified, the quality control accuracy of the time-resolved fluorescence immunoassay analyzer can be improved, in addition, the checking accuracy and the service life of the quality control card are obviously improved through the combined optimization of the factors of preparing the quality control line solution, scribing and sealing the NC film, and the like, and the remarkable economic benefit and application value can be generated.
Drawings
The invention will be described in more detail with reference to the following drawings and specific embodiments:
FIG. 1 is a schematic structural diagram of an electronic quality control card according to an embodiment of the present invention;
fig. 2 is a schematic perspective view of the storage box according to the first embodiment of the invention;
fig. 3 is a schematic view of an internal structure of the storage box according to the first embodiment of the present invention;
FIG. 4 is a graph of mean data and mean trend for the test of example 2.1 of the present invention;
FIG. 5 is a graph of mean data and mean trend for the test of example 2.2.1 in accordance with the present invention;
FIG. 6 is a graph of the test data and the mean trend of the test data of example 2.2.2 in accordance with the present invention;
FIG. 7 is the data of the mean value and the trend chart of the mean value in the laboratory according to the comparative example of the present invention;
FIG. 8 is a graph showing the data and the mean trend of the comparative example of the present invention in a constant temperature and humidity experimental box.
In FIGS. 4-5 and 7, □ represents the mean fluorescence data measured in the first decade of the month, Δ represents the mean fluorescence data measured in the second decade of the month, X represents the mean fluorescence data measured in the third decade of the month, □ represents the first set of fluorescence data measured in the week, △ represents the second set of fluorescence data measured in the week, and X represents the third set of fluorescence data measured in the week in FIGS. 6 and 8.
In the figure: 1-box body; 2-pores; 3-a card slot; 4-electronic quality control card; and 5-a dehumidification groove.
Detailed Description
The technical solutions of the present invention will be described clearly and completely with reference to the following embodiments, 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.
Example one
The embodiment relates to an electronic quality control card for a time-resolved fluorescence immunoassay analyzer, which comprises a reagent shell and a quality control NC membrane, wherein the quality control NC membrane is loaded with two fluorescence quality control lines and is prepared by the following steps:
a. obtaining a fluorescent substance line control solution;
diluting carboxyl modified time-resolved fluorescent microspheres step by step according to high and low concentrations by using 45mmol/L PB solution containing 0.01% (mass percentage content) SDS, and performing microscopic examination after dilution, wherein the fluorescent microspheres are not crosslinked and are in a single state, so as to obtain one quality control line solution with high fluorescence intensity and the other quality control line solution with low fluorescence intensity; wherein the material of the time-resolved fluorescent microspheres is polystyrene;
b. marking a fluorescence quality control line on the NC film;
the film scratching and drying can be completed by adopting a step method:
b1 scratching film
B, placing the quality control line solution obtained in the step a on a film scribing instrument, simultaneously placing an NC film roll on a reel, then scribing or spraying two quality control line solutions with high fluorescence intensity and low fluorescence intensity on different positions of the surface of the NC film through the film scribing instrument with a plurality of film scribing nozzles, and obtaining a fluorescence high-value quality control line and a fluorescence low-value quality control line required by the time-resolved fluorescence immunoassay analyzer;
b2, scribing and drying:
after finishing scribing the membrane, winding the NC membrane, and then placing the wound NC membrane in a 50 ℃ oven for fully drying;
or finishing film scratching and drying by adopting a continuous method:
b, placing the quality control line solution obtained in the step a on a film scribing instrument, simultaneously placing an NC film roll on a reel, then scribing or spraying two quality control line solutions with high fluorescence intensity and low fluorescence intensity on different positions of the surface of the NC film through the film scribing instrument with a plurality of film scribing nozzles, and obtaining a fluorescence high-value quality control line and a fluorescence low-value quality control line required by the time-resolved fluorescence immunoassay analyzer; simultaneously scribing the rear end of the NC film, immediately entering the front end of the scribed NC film into a drying tower preheated to the drying temperature of 30-35 ℃ in advance, namely, continuously operating the processes of scribing the film and drying the film after scribing, and drying the NC film in the drying tower; the running time of the fluorescence quality control line on the NC film from the beginning of scribing to the completion of drying is controlled to be 6 min;
c. sealing and drying the membrane;
the membrane sealing and drying can be completed by adopting a step-by-step method:
c1, film sealing
C, placing the NC membrane dried in the step b2 on a reel again, and then enabling the NC membrane to pass through a soaking solution tank filled with sealing solution at a constant speed to seal the NC membrane;
the confining liquid mainly comprises 1.0 mass percent of PVA with the polymerization degree of 9000, 0.6 mass percent of SDS and 98.4 mass percent of PB solution with the pH value of 7.0-8.0 and 50-60 mmol/L.
c2 drying after sealing
After sealing is finished, winding the NC membrane without cleaning, and then placing the wound NC membrane in a 50 ℃ oven for fully drying to obtain a quality control NC membrane;
or adopting a continuous method to complete membrane sealing and drying:
c, placing the NC membrane dried in the step b2 on a reel again, and then enabling the NC membrane to pass through a soaking solution tank filled with sealing solution at a constant speed to seal the NC membrane; sealing the rear end of the NC membrane, and simultaneously, immediately entering the front end of the sealed NC membrane into a drying tower preheated to the drying temperature of 30-35 ℃ in advance, namely, continuously operating the membrane sealing and drying processes after sealing, and rolling after drying is finished, so as to obtain a quality control NC membrane; the running time of the NC membrane from the beginning of sealing to the completion of drying is controlled to be 6 min;
the confining liquid mainly comprises 1.0 mass percent of PVA with the polymerization degree of 9000, 0.6 mass percent of SDS and 98.4 mass percent of PB solution with the pH value of 7.0-8.0 and 50-60 mmol/L.
The quality control NC film obtained in the first embodiment is cut into strips, and assembled onto a reagent case dedicated for a conventional electronic quality control card to form an electronic quality control card, as shown in fig. 1, in order to facilitate standard comparison of quality control data, a two-dimensional code pattern is further attached to the reagent case, which is advantageous for quality control verification.
In order to better store the electronic quality control card, the invention also designs a storage box suitable for storing the quality control card, as shown in fig. 2 and 3, the storage box has a structure that the storage box comprises a box body 1 with a hollow inner part, sponge or foam which plays a role of shock resistance is filled in the box body 1, a sealable cover body is arranged at the top of the box body 1, an air hole 2 with a one-way sealing valve is arranged on the cover body, and a clamping groove 3 for storing the electronic quality control card 4 and a dehumidification groove 5 for storing a drying agent are arranged in the box body 1.
Example two
The embodiment relates to the evaluation of the electronic quality control card on the verification result of a time-resolved fluorescence immunoassay analyzer.
Example 2.1
This example relates to the comparison of time-resolved fluoroimmunoassay calibration with conventional calibration using the electronic quality control card of the present invention.
The operation process of the electronic quality control card checking time-resolved fluorescence immunoassay analyzer comprises the following steps:
the first use of the quality control card requires inputting information on a matched RFID card to a time-resolved fluorescence analyzer; the quality control card is inserted into the analyzer, the analyzer firstly identifies the two-dimensional code on the quality control card, the identification is completed, the analyzer stores the parameter data of the quality control card, the analyzer automatically scans the fluorescence high-value and fluorescence low-value quality control lines on the quality control card, and compares the scanning result with the parameter data matched with the quality control card to judge whether the quality control is qualified. The above operations were performed under laboratory conditions, and an electronic quality control card was stored in a sealed box.
The operation process of the conventional verification time-resolved fluoroimmunoassay analyzer is as follows:
inputting the information of the matched RFID cards of the fluorescent high-value electronic quality control card and the fluorescent low-value electronic quality control card into an analyzer, inserting the two quality control cards into the analyzer twice, identifying the two-dimensional code on the quality control card by the analyzer, completing identification, storing parameter data of the quality control card in the analyzer, automatically scanning a fluorescent high-value or fluorescent low-value quality control line on the quality control card by the analyzer, comparing the scanning result with the parameter data matched with the quality control card, and judging whether the quality control is qualified. The above procedure was performed under laboratory conditions and the two fluorescent control cards were stored in two conventional storage boxes.
The fluorescence setting value of the high-value fluorescence quality control line is 550000, the fluorescence setting value of the low-value fluorescence quality control line is 55000, the fluorescence values of the two fluorescence quality control lines on the electronic quality control card are tested, the test time is 12 months, the test is carried out 3 times per month respectively in the upper, middle and lower ten days of each month, the test result of each time is obtained by averaging 3 groups of data of the test, each fluorescence line is tested for 36 times in total, the test data of each month are averaged, the change trend of continuous test is obtained, and the result is shown in the following table 1:
the data and the mean trend of the test in each month are shown in fig. 4, the uniformity of the 36 times of the test is analyzed, and the CV value of the fluorescence value of the quality control line is calculated, and the results are shown in table 2 below:
the smaller the SD value and the CV value, the more accurate and stable the quality control result. When the time-resolved fluoroimmunoassay analyzer is verified by adopting two quality control cards, the verification times are increased, and the two quality control cards need to be stored separately; however, it is difficult to ensure that the two quality control cards have the same storage conditions due to environmental problems, and if one of the two electronic quality control cards has a problem, the quality control card fails to be checked, and it is impossible to directly determine whether a problem occurs in a system inside the time-resolved fluorescence analyzer or whether the electronic quality control card is attenuated.
Therefore, the quality control card not only simplifies the verification process, but also eliminates the error caused by two times of verification operations of low-value quality control and high-value quality control which are respectively carried out by two quality control cards in the conventional operation, and obviously improves the quality control accuracy of the time-resolved fluorescence immunoassay analyzer.
Example 2.2
This embodiment relates to the receiver to the influence of electron quality control card check-up precision.
Example 2.2.1
Under the conventional laboratory conditions, the electronic quality control card prepared by continuous film scribing, drying combined with continuous sealing and drying in the first embodiment is placed in the dry sealed storage box (hereinafter, the electronic quality control card assembly in the first embodiment is abbreviated as the first embodiment) of the invention, and the electronic quality control card prepared by continuous film scribing, drying combined with continuous sealing and drying in the first embodiment is placed in a common box (hereinafter, the electronic quality control card in the first embodiment and the common box are abbreviated as the first embodiment) is compared.
The test was performed in the same manner as in example 2.1, and the fluorescence values of the two fluorescence quality control lines on the electronic quality control card were measured, wherein the fluorescence setting value of the high fluorescence quality control line was 550000, the fluorescence setting value of the low fluorescence quality control line was 55000, the test time was 12 months, the test was performed 3 times per month, respectively in the upper, middle and lower ten days of each month, the test results were averaged from the 3 sets of data tested, each fluorescence line was tested 36 times in total, and the test data of each month were averaged to obtain the variation trend of the continuous test, as shown in table 3 below:
the data and the mean trend of the test data per month in the conventional laboratory are shown in fig. 5, the uniformity of the 36 times of the test data is analyzed, and the CV value of the fluorescence value of the quality control line is calculated, and the results are shown in the following table 4:
example 2.2.2
Under the condition of a constant temperature and humidity experimental box, the electronic quality control card prepared by continuous film scratching, drying and continuous sealing and drying in the first embodiment is placed in the dry sealed containing box (the first electronic quality control card component is abbreviated as the first embodiment in the subsequent table), and the electronic quality control card prepared by continuous film scratching, drying and continuous sealing is placed in a common box (the first electronic quality control card and the common box are abbreviated as the second embodiment in the subsequent table) for comparison.
The test was performed in the same manner as in example 2.1, and the fluorescence values of the two fluorescence control lines on the electronic quality control card were measured, wherein the fluorescence setting value of the high fluorescence control line was 550000, the fluorescence setting value of the low fluorescence control line was 55000, the temperature in the experimental box was 37 ℃, the relative humidity was 70%, the illumination throughout the day, the fluorescent lamp power was 30W, the test time was 5 weeks, the test was performed once every 7 days, 3 groups of data were tested each time, the mean value of the fluorescence values tested each week was obtained from the mean value of the 3 groups of data tested each time, and the variation trend of the continuous test was obtained from the mean value of the fluorescence values, as shown in table 5 below:
the data trend of the test under the constant temperature and humidity experimental box is shown in the attached figure 6, 15 groups of data tested are analyzed for homogeneity, and the CV value of the fluorescence value of the quality control line is calculated, and the results are shown in the following table 6:
the results of the two embodiments show that the dry sealed storage box of the invention can also significantly reduce the attenuation influence of the external environment on the fluorescence intensity of the quality control card, so that the stability and the use effectiveness of the quality control card are maintained.
Comparative example
The comparative example relates to the verification performance of quality control cards prepared under different conditions.
Comparative example 1
Common methods treat NC membranes: treating the dried NC membrane with a high-value and low-value quality control line by using a conventional treatment fluid (the conventional treatment fluid contains a hydrophilic surfactant, namely the conventional treatment fluid consists of 0.6 mass percent of Tween 20 and 99.4 mass percent of PB solution with the pH value of 7.0-8.0 and 50-60 mmol/L, aiming at improving the hydrophilic capacity of the NC membrane and helping a fluorescent microsphere solution to be adsorbed on the NC membrane), wherein the NC membrane is not closed and then dried, and the obtained quality control NC membrane is prepared into a quality control card; wherein the run time and temperature for NC film drying were the same as in example 1.
Comparative example 2
Gelatin modified NC membrane: and (3) spraying gelatin on the surface of the NC film with the high-value and low-value quality control lines and drying, and fully drying to prepare the quality control card.
The quality control cards of comparative example 1 were tested for proof performance results under conventional laboratory conditions in a case where the fluorescent quality control cards were conventionally placed (i.e., comparative example 1.1), in a case where the quality control cards of comparative example 1 were placed in the storage case of the present invention (i.e., comparative example 1.2), and in a case where the quality control cards of comparative example 2 were placed in the cases conventionally placed in the fluorescent quality control cards (i.e., comparative example 2.1).
The test was performed in the same manner as in example 2.1, and the fluorescence values of the two fluorescence quality control lines on the electronic quality control card were measured, wherein the fluorescence setting value of the high fluorescence quality control line was 550000, the fluorescence setting value of the low fluorescence quality control line was 55000, the test time was 12 months, 3 times per month were measured in the upper, middle and lower ten days of each month, respectively, the test results were averaged from the 3 sets of data tested, each fluorescence line was tested 36 times in total, and the test data of each month were averaged to obtain the variation trend of the continuous test, as shown in table 7 below:
the data and the mean trend of the test data per month in the conventional laboratory are shown in fig. 7, the uniformity of the 36 times of the test data is analyzed, and the CV value of the fluorescence value of the quality control line is calculated, and the results are shown in the following table 8:
the quality control cards of comparative example 1 were tested for the performance results of the verification that the quality control card of comparative example 1 was stored in the cassette conventionally used for placing the fluorescent quality control card (i.e., comparative example 1.1), the quality control card of comparative example 1 was stored in the storage cassette of the present invention (i.e., comparative example 1.2), and the quality control card of comparative example 2 was stored in the cassette conventionally used for placing the fluorescent quality control card (i.e., comparative example 2.1) under the test conditions of the constant temperature and humidity experimental box.
The test was performed according to the procedure of example 2.1, the fluorescence values of the two fluorescence control lines on the electronic quality control card were tested, the fluorescence setting value of the high fluorescence control line was 550000, the fluorescence setting value of the low fluorescence control line was 55000, the temperature in the experimental box was 37 ℃, the relative humidity was 70%, the illumination was performed all day long, the power of the fluorescent lamp was 30W, the test time was 5 weeks, the test was performed once every 7 days, 3 groups of data were tested each time, the mean value of the fluorescence values tested each week was obtained from the mean value of the 3 groups of data tested each time, and the variation trend of the continuous test was obtained from the mean value of the fluorescence values, as shown in table 9 below:
the trend of the data measured in the constant temperature and humidity experimental box per week is shown in fig. 8, the uniformity of 15 groups of data measured is analyzed, and the CV value of the fluorescence value of the quality control line is calculated, and the results are shown in the following table 10:
compared with the experimental results (tables 8 and 10) of the second embodiment (tables 4 and 6), the electronic quality control card prepared by the invention can play an isolation protection role on the fluorescent quality control line by changing the formula of the sealing liquid, resist the influence of the external environment on the fluorescent intensity of the quality control line, and reduce the sensitivity and attenuation on humidity, temperature and illumination, thereby obviously prolonging the service life of the electronic quality control card and ensuring the verification accuracy.
In conclusion, the electronic quality control card is convenient to use, can realize simultaneous verification of the low value detection and the high value detection of the time-resolved fluoroimmunoassay analyzer by one-step quality control operation, can effectively improve the verification accuracy, has longer service life, is not easy to cause fluorescence attenuation due to the influence of external environment, and has remarkable optimization value.
Claims (9)
1. An electronic quality control card for a time-resolved fluorescence immunoassay analyzer is characterized in that: the electronic quality control card comprises a quality control NC film, and two fluorescent quality control lines are loaded on the quality control NC film;
the quality control NC membrane is prepared by the following steps:
a. obtaining a quality control line solution;
diluting the fluorescent microspheres step by using a diluent, and performing microscopic examination after dilution to ensure that the fluorescent microspheres are not crosslinked and are in a single state to obtain fluorescent microsphere solutions with high fluorescence intensity and low fluorescence intensity respectively, namely two quality control line solutions;
the diluent is phosphate buffer solution containing 0.01-0.05% of sodium dodecyl sulfate by mass, wherein the mass concentration of substances in the phosphate buffer solution is 45 mmol/L; the fluorescent microspheres are hydrophilic time-resolved fluorescent microspheres;
b. marking a fluorescence quality control line on the NC film;
placing the quality control line solution obtained in the step a on a film scribing instrument, scribing or spraying two quality control line solutions on different positions of the surface of the NC film by the film scribing instrument to obtain a fluorescence high-value quality control line and a fluorescence low-value quality control line required by a time-resolved fluorescence immunoassay analyzer, and then drying the NC film in a drying tower;
c. sealing and drying the membrane;
c, sealing the NC membrane dried in the step b by using a sealing liquid, and drying the sealed NC membrane again to obtain a quality control NC membrane;
the sealing liquid mainly comprises a sealing component, a surfactant component and a phosphate buffer solution with the pH value of 7.0-8.0 and 50-60 mmol/L, wherein the sealing component is polyvinyl alcohol 9000, the mass percent of the sealing component in the sealing liquid is 1.0-1.3%, the surfactant component is at least one of Pluronic F68, RHODASURF ON-870 and TRITON X-305, and the mass percent of the surfactant component in the sealing liquid is 0.5-1.0%.
2. The electronic quality control card for time-resolved fluoroimmunoassay analyzer according to claim 1, wherein: the electronic quality control card also comprises a reagent shell for assembling the strip-shaped quality control NC membrane, and a two-dimensional code pattern containing quality control standard information is sprayed on the reagent shell.
3. The electronic quality control card for time-resolved fluoroimmunoassay analyzer according to claim 1, wherein: the hydrophilic time-resolved fluorescent microsphere is any one of a carboxyl modified time-resolved fluorescent microsphere, a hydroxyl modified time-resolved fluorescent microsphere, a coupled protein A time-resolved fluorescent microsphere and a coupled protein G time-resolved fluorescent microsphere.
4. The electronic quality control card for time-resolved fluoroimmunoassay analyzer according to claim 1, wherein: b, the operation of scribing or spraying the fluorescent quality control line on the NC film and the subsequent drying process are operated in a continuous state;
the step of continuously operating comprises:
placing the NC film roll on a take-out reel, and then marking or spraying two quality control line solutions diluted to the required concentration on the NC film simultaneously by a film marking instrument; and simultaneously scribing the rear end of the NC film, immediately feeding the front end of the scribed NC film into a drying tower preheated to the drying temperature in advance, and drying.
5. The electronic quality control card for time-resolved fluoroimmunoassay analyzer according to claim 4, wherein: the running time of each fluorescence quality control line on the NC membrane from the beginning of scribing to the completion of drying is 5-6 min, and the drying temperature is 30-35 ℃.
6. The electronic quality control card for time-resolved fluoroimmunoassay analyzer according to claim 1, wherein: c, the operation of membrane sealing and the subsequent drying process are operated in a continuous state;
the step of continuously operating comprises:
and c, enabling the NC membrane dried in the step b to pass through a soaking liquid tank with sealing liquid at a constant speed to seal the NC membrane, sealing the rear end of the NC membrane, simultaneously enabling the front end of the sealed NC membrane to immediately enter a drying tower preheated to the drying temperature in advance, and rolling after drying.
7. The electronic quality control card for time-resolved fluoroimmunoassay analyzer according to claim 6, wherein: the running time of the NC membrane from the beginning of sealing to the completion of drying is 5-6 min, and the drying temperature is 30-35 ℃.
8. The utility model provides a time-resolved fluorescence immunoassay appearance is with electronic quality accuse card subassembly which characterized in that: comprising an electronic quality control card according to any one of claims 1 to 7, and a storage case for storing the electronic quality control card; the storage box comprises a box body with a hollow interior, a sealable cover body is arranged at the top of the box body, an air hole with a one-way sealing valve is formed in the cover body, and a clamping groove for containing the electronic quality control card and a dehumidification groove for storing a drying agent are formed in the box body.
9. The electronic quality control card assembly for time-resolved fluoroimmunoassay analyzer of claim 8, wherein: the coefficient of variation value of the electronic quality control card is not more than 0.4% in one year.
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