CN107664695B - Calibration sheet and calibration method for dry-type immunoassay analyzer - Google Patents
Calibration sheet and calibration method for dry-type immunoassay analyzer Download PDFInfo
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Abstract
本发明公开了一种用于干式免疫分析仪校准的校准片及校准方法，该校准片由上盖板和下盖板盖合构成，该上盖板上设置有进样窗和出气孔，该下盖板设置有沟道，该沟道至少一端具有容纳腔，其中，下盖板的沟道的一个容纳腔与上盖板的进样窗在组合安装时位置上下对应。本发明的校准片具有不同形状和加工深度的沟道，通过加入不同的液体染料及染料浓度，可实现干式免疫分析仪自动加样评估、自动稀释评估、定量检测等功能的调试校准。
The present invention discloses a calibration sheet and a calibration method for calibrating a dry immunoassay analyzer. The calibration sheet is composed of an upper cover plate and a lower cover plate, wherein the upper cover plate is provided with a sample injection window and an air outlet, and the lower cover plate is provided with a channel, wherein at least one end of the channel has a receiving cavity, wherein a receiving cavity of the channel of the lower cover plate corresponds to the sample injection window of the upper cover plate in the upper and lower positions when assembled and installed. The calibration sheet of the present invention has channels of different shapes and processing depths, and by adding different liquid dyes and dye concentrations, the debugging and calibration of the functions of the dry immunoassay analyzer such as automatic sample addition evaluation, automatic dilution evaluation, and quantitative detection can be realized.
Description
Technical Field
The invention belongs to the technical field of instrument calibration, and relates to a calibration sheet which is specially designed, and can be used for calibration evaluation of a dry-type immunoassay analyzer by adding specific dyes.
Background
The dry immunoassay analyzer is used as POCT (point-of-CARE TESTING) equipment and is widely applied to the field of biomedical inspection. The labeling substances that can be used for quantitative analysis of analytes are varied from colloidal gold, colored latex microspheres, to fluorescent microspheres, quantum dots, and the like. However, when actually calibrating the device, we find the following problems: 1. the calibration sheets used for calibration at present are mostly dry sheets, and cannot be used for liquid path system evaluation, dilution evaluation and whole system evaluation; 2. the calibration sheet used for calibration has instability, such as a fluorescent dry sheet and a quantum dot dry sheet; 3. calibration tiles used for calibration also suffer from serious non-uniformity problems, such as gray-scale tiles for inkjet printing. The above problems create a significant disturbance in the overall calibration and standardized analysis of the dry immunoassay analyzer.
Conventional cuvettes for use in fluid path systems are difficult to meet the requirements of ultra-thin calibration sheets in thickness required for dry immunoassays. The appearance thickness of the calibration piece is 3.2-3.7 mm, and can be flexibly designed according to different requirements in practice, but the cuvette is of a fixed structure, the appearance thickness is about 1cm, and if the cuvette with the minimum volume is used, the appearance thickness is unchanged. Furthermore, even with the minimum specification of cuvette, the design of the lumen on the order of μm is less likely to occur.
Based on the reasons and requirements, the calibration piece with the special flow channel is designed, and different dyes and dye concentrations can be added according to the requirements.
Disclosure of Invention
The invention aims to provide a calibration sheet for calibrating a dry immunoassay analyzer, which solves the problems of instability and non-uniformity of the existing dry calibration sheet.
In order to achieve the above purpose, the invention provides a calibration piece for calibrating a dry-type immunoassay analyzer, which is formed by covering an upper cover plate and a lower cover plate, wherein the upper cover plate is provided with a sample inlet window and an air outlet hole, the lower cover plate is provided with a channel, at least one end of the channel is provided with a containing cavity, and one containing cavity of the channel of the lower cover plate corresponds to the sample inlet window of the upper cover plate in a vertical position during combined installation.
Preferably, the channels are in the shape of a plurality of continuous U-shapes.
Preferably, the two ends of the channel are provided with accommodating cavities.
Preferably, the U-shaped is uniformly arranged, the U-shaped openings are uniform in size, and one end of the channel is provided with a containing cavity.
Preferably, the channel is in a straight line shape, and the two ends of the channel are provided with accommodating cavities.
Preferably, the channels form a matrix array by a plurality of rectangular holes with volumes gradually increasing or decreasing from left to right, and a central axis channel penetrates through the whole array and is connected with the accommodating cavities at two ends. The rectangular array holes are not limited to rectangular, but may be elliptical or circular, etc. The array features a series of holes with increasing or decreasing volumes and a channel for connecting the holes, the sample inlet window and the waste liquid pool (or called liquid storage pool), so that one sample can be added at the sample inlet window, and signals with different intensities can be generated in the array region (the generation of the signals is not only related to the concentration of the solution, but also related to the thickness and the sectional area of the liquid penetrated by the light path).
Preferably, the upper cover plate and the lower cover plate are connected through a non-connection or one end.
The invention also provides a using method of the calibration sheet for calibrating the dry-type immunoassay analyzer, which comprises the following steps:
Step 1, adding a liquid sample to be measured into a sample inlet window of a calibration sheet;
Step 2, scanning the calibration sheet by adopting a detection module of a standardized dry immunoassay analyzer to obtain a standard signal;
Step 3, placing the calibration sheet in a detection module of a dry immunoassay analyzer to be detected for scanning, and collecting signals;
and 4, comparing the acquired signal with a standard signal to calibrate a detection module of the dry immunoassay analyzer to be detected.
A series of micro flow channels (namely channels) are designed on the surface of the lower cover plate, and holes are punched on the upper cover plate and serve as sample inlet windows and air outlet holes. The upper cover plate and the lower cover plate are processed by bonding, bonding or buckling to form a calibration sheet for calibration. Liquid dye is added at the sample adding window of the flow channel, and driven by capillary action, the liquid dye can infiltrate downwards and migrate to the front section, and the dye can submerge the whole flow channel and reach an equilibrium state within 1 minute. The shape and thickness of the channel determines the volume of liquid (channel area: channel depth) that can be accommodated, for example, a channel with a thickness of 50 μm can accommodate 15 μl of sample, and a channel with a thickness of 1mm can accommodate 300 μl of sample. In addition, the thickness of the flow channel is increased, the corresponding optical signal is changed, and the signal rises (emitted or reflected light detection) or falls (absorbed light detection) according to the detection principle.
The flow channels have different shapes and combinations according to the evaluation content of the equipment to be tested, and generally comprise snakes, matrix columns, linearity, holes and the like.
The invention has the advantages that:
1. The calibration sheet can add a specified amount of specific dyes with different excitation emission spectrums at the sample adding window according to the equipment calibration requirement, realize the detection of the optical module under the spectrum condition, and calibrate the light source detection module according to the difference between the detection signal and the standard value. If the visible light detection module is detected and calibrated, conventional dyes such as ponceau and the like can be added; when evaluating the fluorescence detection module, a specific fluorescent dye should be added.
2. The calibration sheet is provided with different flow channels, and when the flow channels deviate in the horizontal direction or the ray direction of the equipment in a combined way, the detected signals deviate from the standard values correspondingly, so that the detection and the calibration of the horizontal direction and the ray direction of the equipment can be carried out.
3. The calibration sheet attempts to design holes with different sizes on the same lower cover plate, the holes are communicated with each other, the dye concentration is consistent, and the hole volume is in an increasing or decreasing trend. Therefore, it is considered that the same sample can generate signals with different intensities at different holes, and the difference between the signals should have consistency with the difference of the hole volumes, if there is a significant difference, the difference should be related to the resolution of the light source detection module, and the ratio of the difference of the signals with high and low values to the difference of the hole volumes is used as the evaluation standard of the resolution of the device, so that the device can be calibrated.
4. The calibration sheet can be used for an optical detection module of evaluation equipment, and can well avoid the problems of fluorescence quenching, attenuation and the like of dye in repeated excitation due to the disposable characteristic of the calibration sheet. Meanwhile, due to the characteristic of solution uniformity, the negative influence on equipment calibration caused by uneven dry plates during ink-jet printing can be well avoided.
5. The calibration sheet provided by the invention can evaluate the automatic dilution precision of equipment according to equipment requirements.
6. The calibration sheet provided by the invention can evaluate the automatic sample adding precision of equipment according to equipment requirements.
Drawings
Fig. 1 is a schematic structural view of a calibration sheet (calibration sheet 1 of example 1) for calibration of a dry immunoassay instrument according to the present invention.
Fig. 2 is a schematic structural view of a calibration sheet (calibration sheet 2 of example 2) for calibration of a dry immunoassay instrument according to the present invention.
Fig. 3 is a schematic structural view of a calibration sheet (calibration sheet 3 of example 3) for calibration of a dry immunoassay instrument according to the present invention.
Fig. 4 is a schematic structural view of a calibration sheet (calibration sheet 4 of example 4) for calibration of a dry immunoassay instrument according to the present invention.
Detailed Description
The technical scheme of the invention is further described below with reference to the accompanying drawings and examples.
Example 1
A calibration sheet for calibrating a dry-type immunoassay analyzer is shown in fig. 1, and is formed by covering an upper cover plate 10 and a lower cover plate 20, wherein the upper cover plate 10 is provided with a sample inlet window 11 and a gas outlet hole 12, and is preferably positioned at two ends of the upper cover plate, the lower cover plate 20 is provided with a channel 21, and two ends of the channel are respectively provided with a containing cavity 211, wherein one containing cavity of the channel corresponds to the sample inlet window 11 of the upper cover plate in a vertical position during combined installation, and the other containing cavity corresponds to the gas outlet hole 12 in a vertical position. The channel is in a plurality of continuous U shapes, and the U-shaped openings are consistent or inconsistent in size.
PMMA (polymethyl methacrylate) is used as a base material, an upper cover plate and a lower cover plate are processed and prepared, a channel with the depth of 50 mu m-1 mm is processed on the lower cover plate, a sample inlet window and a gas outlet hole are punched on the upper cover plate, and then the upper cover plate and the lower cover plate are bonded and fixed or fastened and fixed, so that the calibration sheet 1 is obtained.
Calibration sheet 1 prepared in this example 1 was used for standardized evaluation of the detection module. A standardized dye (such as ponceau red and bromobenzyl violet) is added to the right side sample adding window of the calibration sheet, a 50 μm thick flow channel can contain 15 μl sample, the same flow channel can contain 300 μl sample after processing 1mm thickness, and after the liquid flow reaches balance (about 1 minute), the sample is placed on a standardized visible light detection module for signal acquisition. And then, placing the calibration sheet on an optical detection module of the equipment to be detected for signal acquisition, evaluating whether the optical detection module of the equipment to be detected meets the expected requirement according to the ascending or descending of the signal, and debugging the components of the equipment to be detected until the signal acquired by the equipment to be detected is consistent with the signal acquired by the standardized optical module.
The signal obtained by the calibration sheet is relatively uniform and smooth, signal burrs (i.e. higher background noise caused by uneven ink jet) which are often generated during dry sheet calibration such as ink jet printing do not appear, and the calibration sheet has good advantages when the signal is standardized.
Example 2
A calibration sheet for calibrating a dry-type immunoassay analyzer is shown in fig. 2, and is formed by covering an upper cover plate 10 and a lower cover plate 20, wherein the upper cover plate 10 is provided with a sample inlet window 11 and a gas outlet hole 12, and is preferably positioned at two ends of the upper cover plate, the lower cover plate 20 is provided with a channel 21, and two ends of the channel are respectively provided with a containing cavity 211, wherein one containing cavity of the channel corresponds to the sample inlet window 11 of the upper cover plate in a vertical position during combined installation, and the other containing cavity corresponds to the gas outlet hole 12 in a vertical position. The channels are in a straight line shape and are uniformly paved (the channel width is consistent).
And (3) processing and preparing an upper cover plate and a lower cover plate by taking PDMS as a base material, processing a channel with the depth of 50 mu m-1 mm on the lower cover plate, punching and preparing a sample inlet window 11 and a gas outlet hole 12 on the upper cover plate, and then bonding and fixing or buckling and fixing the upper cover plate and the lower cover plate to obtain the calibration sheet 2.
The calibration sheets prepared in example 1 and example 2 were used in combination for evaluation of the horizontal and vertical directions of the apparatus. Adding a certain amount of standardized solution into the right sample adding windows of the calibration sheet 1 and the calibration sheet 2, after balancing, placing the solutions into a fluorescence detection module of equipment to be detected, scanning from right to left, and generating signals, and judging that the horizontal calibration of the equipment is problematic if the signals of the calibration sheet 1 and the calibration sheet 2 are simultaneously lifted or lowered on one side; only the calibration patch 2 is signal deviated, indicating that the ray direction is not completely calibrated. Because the calibration sheet belongs to disposable consumable, the added fluorescent substance is only excited for 1 time and is discarded, and the defect that the fluorescent substance is unstable due to repeated excitation is well avoided.
It can be seen that the use of both calibration sheet 1 and calibration sheet 2 can be used to evaluate the calibration of the device in both the horizontal and radial directions.
Example 3
A calibration sheet for calibrating a dry-type immunoassay analyzer is shown in fig. 3, and is formed by covering an upper cover plate 10 and a lower cover plate 20, wherein the upper cover plate 10 is provided with a sample inlet window 11 and a gas outlet hole 12, and is preferably positioned at two ends of the upper cover plate, the lower cover plate 20 is provided with a channel 21, and two ends of the channel are respectively provided with a containing cavity 211, wherein one containing cavity of the channel corresponds to the sample inlet window 11 of the upper cover plate in a vertical position during combined installation, and the other containing cavity corresponds to the gas outlet hole 12 in a vertical position. The channel is composed of a first channel 212 along the longitudinal direction of the lower cover plate and a plurality of second channels 213 which are vertical to the first channel and are communicated with the first channel, the first channels 212 are in a straight line shape, the second channels 213 are uniformly arranged at intervals, each second channel 213 is divided into two symmetrical sections by the first channel 212, and the lengths of the second channels are positioned at two sides of the first channel 212 and are in an increasing or decreasing trend. Preferably, the length of the second channel decreases from the receiving chamber corresponding to the loading window to the receiving chamber on the other side (corresponding to the loading hole).
And (3) processing and preparing an upper cover plate and a lower cover plate by taking BF33 high borosilicate glass as a base material, processing a channel with the depth of 50 mu m-1 mm on the lower cover plate, punching and preparing a sample inlet window 11 and a gas outlet hole 12 on the upper cover plate, and then carrying out bonding fixation or buckling fixation on the upper cover plate and the lower cover plate to obtain the calibration sheet 3.
The calibration sheet 3 is used for evaluation of the detectable range and resolution. The right sample adding window of the calibration sheet 3 is added with specific dyes such as ponceau or fluorescein, which can be used for evaluating and calibrating the resolution and detection range of the dry type immunization equipment, the signals from left to right should be increased in sequence, the resolution and the detection range of the equipment can be evaluated according to the signal amplification difference and the signal value at the lowest position, and the equipment can be ensured to have better resolution when reaching a certain response, thereby meeting the requirement of effectively standardizing different equipment.
Example 4
As shown in fig. 4, the calibration sheet for calibrating the dry-type immunoassay analyzer is formed by covering an upper cover plate 10 and a lower cover plate 20, wherein the upper cover plate 10 is provided with a sample inlet window 11 and a gas outlet hole 12, preferably positioned at two ends of the upper cover plate, the lower cover plate 20 is provided with a channel 21, one end of the channel is provided with a containing cavity 211, and the containing cavity corresponds to the sample inlet window 11 of the upper cover plate in a vertical position during combined installation. The channels are in a plurality of continuous U shapes, and the U-shaped openings are uniform in size and are uniformly paved.
And (3) processing and preparing an upper cover plate and a lower cover plate by taking quartz glass as a base material, processing a channel with the depth of 200 mu m on the lower cover plate, punching and preparing a sample inlet window 11 and a gas outlet hole 12 on the upper cover plate, and then bonding and fixing or buckling and fixing the upper cover plate and the lower cover plate to obtain the calibration sheet 4.
The calibration sheet 4 is used for evaluation of the automatic loading accuracy. The channel of the calibration sheet 4 needs to precisely control the machining error. If the device is designed to hold 70 mu L of sample, the device can be used for evaluating the accuracy of the sample loading within 60 mu L of the automatic loading dry immunoassay analyzer, and can be used for judging the sample loading accuracy of the device according to the length of a sample filling channel and the continuous peak area of the signal acquired on a standardized detection module.
Example 5
The calibration sheet 1 prepared in example 1 can also be used for evaluation of the accuracy of automatic dilution. The accuracy of the autodilution can be determined by using a standardized mother liquor of 100 x (i.e., 100 times, if the sample is 1g/mL, 100 x mother liquor is 100g/mL; if the sample is 0.01%,100 x mother liquor is 1%, if the sample is 1mmol/L,100 x mother liquor is 100 mmol/L), automatically diluting the standardized mother liquor into 1x sample (i.e., diluting 100 times) by equipment, adding the 1x sample into a sample adding window on the right side of the calibration sheet 1, collecting corresponding signals on the optical detection module, and comparing the signals generated by the 1x standardized solution.
Example 6
The calibration sheet provided by the invention can evaluate the automatic sample adding precision of equipment according to equipment requirements.
And (3) using the standardized 1X solution, automatically sampling by using equipment to be detected, dripping the sample onto a calibration sheet (through a sample adding window), detecting by using standardized detection equipment, collecting optical detection signals, and comparing the collected optical detection signals with signals detected by the standardized 1X solution by using the standardized detection equipment to judge the sample adding precision.
While the present invention has been described in detail through the foregoing description of the preferred embodiment, it should be understood that the foregoing description is not to be considered as limiting the invention. Many modifications and substitutions of the present invention will become apparent to those of ordinary skill in the art upon reading the foregoing. Accordingly, the scope of the invention should be limited only by the attached claims.
Claims (4)
1. The calibration sheet is characterized by comprising an upper cover plate and a lower cover plate which are covered, wherein the upper cover plate is provided with a sample inlet window and a gas outlet hole, the lower cover plate is provided with a channel, two ends of the channel are respectively provided with a containing cavity, and one containing cavity of the channel of the lower cover plate corresponds to the sample inlet window of the upper cover plate in a combined installation mode; the channel is formed into a matrix array by a plurality of rectangular holes with sequentially increasing or decreasing volumes from left to right, and a central axis channel penetrates through the whole matrix array and is connected with the accommodating cavities at two ends.
2. The calibration sheet for calibrating a dry-type immunoassay analyzer of claim 1, wherein the upper cover plate and the lower cover plate are connected by a non-connection or one end.
3. The calibration sheet for calibrating a dry-type immunoassay analyzer of claim 1, wherein the upper cover plate and the lower cover plate are bonded or snap-fit.
4. A method of using the calibration patch for calibration of a dry immunoassay according to claim 1, the method comprising the steps of:
Step 1, adding a liquid sample to be measured into a sample inlet window of a calibration sheet;
Step 2, scanning the calibration sheet by adopting a detection module of a standardized dry immunoassay analyzer to obtain a standard signal;
Step 3, placing the calibration sheet in a detection module of a dry immunoassay analyzer to be detected for scanning, and collecting signals;
and 4, comparing the acquired signal with a standard signal to calibrate a detection module of the dry immunoassay analyzer to be detected.
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| CN108535239B (en) * | 2018-03-28 | 2021-05-25 | 上海艾瑞德生物科技有限公司 | Micro-fluidic chip and detection system based on micro-droplets |
| CN109444118B (en) * | 2018-12-24 | 2021-07-27 | 三诺生物传感股份有限公司 | Measuring method and device for chemical luminous inspection instrument |
| CN113252633B (en) * | 2021-07-14 | 2021-10-12 | 季华实验室 | A kind of quality control detection method and standard plate of liquid phase chip analyzer |
| DE102023121083A1 (en) * | 2023-08-08 | 2025-02-13 | Orphan Diagnostics AS | Fluid-storing test carrier channel structure and microfluidic test carrier |
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| CN2695954Y (en) * | 2004-04-27 | 2005-04-27 | 中国兵器工业集团第五三研究所 | Standard plate investigating / correcting for enzymic standard analyzer |
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