CN111650170B - Metering standard for calibrating luminous immunity analyzer for bottom or side direction detection - Google Patents
Metering standard for calibrating luminous immunity analyzer for bottom or side direction detectionInfo
- Publication number
- CN111650170B CN111650170B CN202010635924.4A CN202010635924A CN111650170B CN 111650170 B CN111650170 B CN 111650170B CN 202010635924 A CN202010635924 A CN 202010635924A CN 111650170 B CN111650170 B CN 111650170B
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- light source
- light
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- filter
- detection
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- 238000001514 detection method Methods 0.000 title claims abstract description 59
- 230000036039 immunity Effects 0.000 title description 3
- 238000003018 immunoassay Methods 0.000 claims abstract description 47
- 230000007935 neutral effect Effects 0.000 claims abstract description 30
- 238000001228 spectrum Methods 0.000 claims abstract description 16
- YZCKVEUIGOORGS-NJFSPNSNSA-N Tritium Chemical compound [3H] YZCKVEUIGOORGS-NJFSPNSNSA-N 0.000 claims description 27
- 238000004020 luminiscence type Methods 0.000 claims description 27
- 229910052722 tritium Inorganic materials 0.000 claims description 27
- 238000005259 measurement Methods 0.000 claims description 15
- 230000003595 spectral effect Effects 0.000 claims description 15
- 238000002834 transmittance Methods 0.000 claims description 10
- 239000012460 protein solution Substances 0.000 claims description 5
- 239000011521 glass Substances 0.000 claims description 4
- 239000010453 quartz Substances 0.000 claims description 4
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N silicon dioxide Inorganic materials O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 4
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- 238000011156 evaluation Methods 0.000 description 5
- 238000002965 ELISA Methods 0.000 description 4
- 239000000047 product Substances 0.000 description 4
- 238000004451 qualitative analysis Methods 0.000 description 4
- 238000004445 quantitative analysis Methods 0.000 description 4
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- 102000004190 Enzymes Human genes 0.000 description 3
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- 238000004458 analytical method Methods 0.000 description 2
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- 238000000338 in vitro Methods 0.000 description 2
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- 238000000034 method Methods 0.000 description 2
- 238000012795 verification Methods 0.000 description 2
- 238000002835 absorbance Methods 0.000 description 1
- 239000000427 antigen Substances 0.000 description 1
- 102000036639 antigens Human genes 0.000 description 1
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- 238000005401 electroluminescence Methods 0.000 description 1
- BHEPBYXIRTUNPN-UHFFFAOYSA-N hydridophosphorus(.) (triplet) Chemical compound [PH] BHEPBYXIRTUNPN-UHFFFAOYSA-N 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 229920002521 macromolecule Polymers 0.000 description 1
- 230000005180 public health Effects 0.000 description 1
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- 239000000126 substance Substances 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
- 150000003649 tritium Chemical class 0.000 description 1
Abstract
The invention relates to a measuring standard for calibrating a luminous immunoassay analyzer for bottom or side detection, which comprises the following components: the light source is fixed in the groove, the cylindrical lateral light guide is fixed at the lower end of the bracket, the cylindrical barrel is a light filter cavity between the light source and the lateral light guide, three pairs of jacks are arranged along the length direction of the light filter cavity, each pair of jacks is arranged at the circumferential symmetrical position of the cylindrical barrel, one or more of the interference filter, the attenuation sheet and the spectrum neutral filter are respectively inserted into the pair of jacks, the interference filter, the attenuation sheet and the spectrum neutral filter are placed in parallel along the length direction of the light filter cavity, and the light outlet hole of the light source is opposite to one or more of the interference filter, the attenuation sheet and the spectrum neutral filter.
Description
Technical Field
The invention relates to the field of biotechnology and in-vitro diagnosis, in particular to a measurement standard device for calibrating a luminous immunoassay analyzer for bottom or lateral detection and application thereof.
Background
Immunoassay is a means of qualitative and quantitative analysis using specific interaction between antigen and antibody, and may include enzyme-linked immunoassay, fluorescent immunoassay, luminescent immunoassay, etc. according to the difference in immunoassay reporting modes. Enzyme-linked immunoassay utilizes enzyme-labeled antibodies to catalyze substrates to generate colored products, and qualitative and quantitative analysis is realized by measuring absorbance or optical density of the products; the fluorescent immunoassay is to utilize enzyme-labeled antibody to catalyze a substrate to generate a product with fluorescence, and the qualitative and quantitative analysis is realized by measuring the fluorescence signal of the product, so that the fluorescent immunoassay has higher sensitivity compared with the enzyme-linked immunoassay; the luminescence immunoassay is to utilize enzyme-labeled antibody to catalyze a substrate to generate chemiluminescence or generate a luminescence signal through an electroluminescence means, and the qualitative and quantitative detection of a target object is realized through detection of the luminescence signal. Compared with enzyme-linked immunoassay and fluorescent immunoassay, the luminescent immunoassay has higher sensitivity due to no influence of a background light source. Currently, luminescence immunoassay is widely used in various fields of medical examination, in-vitro diagnosis and the like, and whether an analysis result is accurate or not is related to public health and safety.
The luminescence immunoassay instrument has various structures, including an open system and a closed system. Open systems can be divided from sample loading systems into plate and tube systems, with closed systems being predominantly tubular. In any structural form, qualitative and quantitative analysis is carried out by detecting the final luminescence signal. From the light-emitting position detected by the detector, the three modes of top detection, bottom detection and lateral detection are mainly included. As the name suggests, top detection is performed above the sample tube, bottom detection is performed below the sample tube, and lateral detection is performed in a lateral orientation of the sample tube. Wherein, bottom detection and lateral detection in the luminescence immunoassay analyzer are main detection modes, and manufacturers using top detection are very few.
The accuracy of the detection result of the luminescence immunoassay is closely related to the metering performance of the optical detection system of the instrument, wherein the metering performance comprises indexes such as accuracy, sensitivity, repeatability and the like, and the confirmation and calibration of the metering performance indexes must be realized through metering standards. Because luminescence immunity is a technology newly developed in recent years, the development of the measurement standard is relatively lagged, and at present, only a top detection measurement standard is adopted in the market, but the main current instrument in the market adopts bottom detection and lateral detection, the existing measurement standard cannot be used for the bottom detection and lateral detection instrument, and the quality of an analysis measurement result of the instrument is difficult to ensure. Therefore, it is urgent to develop a measuring standard for a luminescence immunoassay analyzer capable of simultaneously performing bottom detection and lateral detection.
Disclosure of Invention
The invention aims to fill the blank of calibrating a metering standard for calibrating a luminous immunoassay analyzer for bottom detection and lateral detection, and provides a metering standard for calibrating the luminous immunoassay analyzer for bottom or lateral detection, which has the advantages of high stability, no maintenance, easy operation and low price, and an application thereof.
In order to achieve the aim of the application, the application adopts the following technical scheme:
The invention relates to a measuring standard for calibrating a luminous immunoassay analyzer for bottom or side detection, which comprises the following components: support, light source, side direction light guide, first interference filter, attenuation piece and spectral neutral filter, wherein: the support is a cylinder, a groove is formed in the upper end of the support, the light source is fixed in the groove, a cylindrical lateral light guide is fixed at the lower end of the support, the cylinder between the light source and the lateral light guide is a light filter cavity, three pairs of jacks are formed in the length direction of the light filter cavity, each pair of jacks are formed at symmetrical positions of the circumference of the cylinder, one or more of the interference filter, the attenuation sheet and the spectrum neutral filter are respectively inserted into the pair of jacks, the jacks are placed in parallel along the length direction of the light filter cavity, and a light outlet of the light source faces one or more of the interference filter, the attenuation sheet and the spectrum neutral filter.
The invention relates to a measuring standard for calibrating a luminous immunoassay analyzer for bottom or side detection, wherein: the lateral light guide is a commercial lateral light guide fiber tube or a glass tube or a quartz tube filled with 1% -10% protein solution.
The invention relates to a measuring standard for calibrating a luminous immunoassay analyzer for bottom or side detection, wherein: the interference filter, the attenuation sheet and the spectrum neutral filter are sequentially inserted into the three pairs of sockets from top to bottom along the length direction of the filter cavity.
The invention relates to a measuring standard for calibrating a luminous immunoassay analyzer for bottom or side detection, wherein: the transmittance of the spectral neutral filter is 50%, 30%, 10%, 3% or 1%.
The invention relates to a measuring standard for calibrating a luminous immunoassay analyzer for bottom or side detection, wherein: the lateral light guide is fixed at the lower end of the holder by means of a number of fixing screws along the circumference of the holder.
The invention relates to a measuring standard for calibrating a luminous immunoassay analyzer for bottom or side detection, wherein: the lateral light guide is fixed at the lower end of the bracket by threads.
The invention relates to a measuring standard for calibrating a luminous immunoassay analyzer for bottom or side detection, wherein: the light source is a tritium light source module, the tritium light source module is composed of a tritium light pipe, a second interference filter, a first collimating lens and a light attenuation sheet which are arranged from top to bottom, and a light outlet hole is formed in the lower end of the tritium light source module.
The invention relates to a measuring standard for calibrating a luminous immunoassay analyzer for bottom or side detection, wherein: the light source is an LED light source module, the LED light source module is composed of a button battery, an adjustable constant current source, an LED light source and a second collimating lens which are arranged from top to bottom, and a light outlet hole is formed in the lower end of the LED light source module.
The measuring standard device for calibrating the luminous immunoassay analyzer for bottom or side detection can carry out bottom detection and side detection, and can correct the measurement result of the luminous immunoassay analyzer so as to ensure the detection quality.
Drawings
FIG. 1 is a schematic forward cross-sectional view of a measurement standard of a bottom or side-detection luminescence immunoassay analyzer of the present invention;
FIG. 2 is a schematic cross-sectional view of FIGS. 1 A-A;
FIG. 3 is a schematic perspective view of a tritium light source module;
FIG. 4 is a schematic perspective view of an LED light source module;
FIG. 5 is a schematic diagram of the detection port and photomultiplier positions on a luminescence immunoassay instrument.
In fig. 1 to 5, reference numeral 1 denotes a bracket; reference numeral 2 is a light source; reference numeral 3 is a lateral light guide; reference numeral 4 is a socket; reference numeral 5 denotes a spectral neutral filter; reference numeral 6 is a fixing screw; reference numeral 7 is a tritium light pipe; reference numeral 8 denotes a second interference filter; reference numeral 9 denotes a first collimating lens; reference numeral 10 denotes a light attenuation sheet; reference numeral 11 denotes a light exit hole; reference numeral 12 is a button cell; reference numeral 13 is an adjustable constant current source 1; reference numeral 14 is an LED light source; reference numeral 15 denotes a second collimator lens; reference numeral 16 is a groove; reference numeral 17 denotes an attenuation sheet; reference numeral 18 denotes a first interference filter; reference numeral 19 denotes a luminescence immunoassay analyzer; reference numeral 20 denotes a detection port; reference numeral 21 denotes a filter chamber; reference numeral 22 denotes a photomultiplier tube.
Detailed Description
As shown in fig. 5, the measuring standard for calibrating the luminescence immunoassay analyzer for bottom or side detection of the present invention is inserted into the detection port 20 of the luminescence immunoassay analyzer 19, and the photomultiplier 22 is located at the bottom or side of the luminescence immunoassay analyzer.
As shown in fig. 1 and 2, the measuring standard for calibration of a luminescence immunoassay analyzer for bottom or lateral detection of the present invention includes: support 1, light source 2, side direction light guide 3, first interference filter 18, attenuation piece 17 and spectral neutral filter 5, its characterized in that: the bracket 1 is a cylindrical barrel, a groove 16 is formed in the upper end of the bracket 1, the light source 2 is fixed in the groove 16, a cylindrical lateral light guide 3 is fixed at the lower end of the bracket 1, the lateral light guide 3 is a commercial lateral light guide fiber tube or a glass tube or a quartz tube filled with 1% -10% protein solution, and the lateral light guide 3 is fixed at the lower end of the bracket 1 through a plurality of fixing screws 6 along the circumference of the bracket 1 or is fixed at the lower end of the bracket 1 through threads. The cylindrical barrel between the light source 2 and the lateral light guide 3 is a light filter chamber 21, three pairs of sockets 4 are arranged along the length direction of the light filter chamber 21, each pair of sockets 4 is arranged at the symmetrical position of the circumference of the cylindrical barrel, the interference filter 18, the attenuation sheet 17 and the spectrum neutral filter 5 are sequentially inserted into one pair of sockets 4 from top to bottom, the sockets are arranged in parallel along the length direction of the light filter chamber 21, the light outlet 11 of the light source 2 is opposite to the interference filter 18, the attenuation sheet 17 and the spectrum neutral filter 5, the spectrum neutral filter 5 with the transmittance of 50%, 30%, 10%, 3% or 1% can be replaced in the detection process,
As shown in fig. 3, the light source 2 is a tritium light source module, which is composed of a tritium light pipe 7, a second interference filter 8, a first collimating lens 9 and a light attenuation sheet 10 which are arranged from top to bottom, and a light outlet 11 is formed at the lower end of the tritium light source module. The tritium light source is solid tritium salt or tritium gas, and tritium is a radioactive element with half-life of about 12 years. During decay, beta rays are released, which excite fluorescent or phosphorous substances to fluoresce or phosphorescence. The tritium light source is not required to be powered by a power supply, and the luminous intensity is not interfered by the power supply state; the half-life period of tritium is long, the luminous intensity is basically unchanged and the stability is high in the process of measuring performance indexes for several hours. Meanwhile, as the power supply is not used for supplying power, the design of the device is simplified, and the maintenance and use cost is reduced. The tritium light source module comprises a tritium light pipe, an interference filter, a collimating lens and a light attenuation sheet, wherein the tritium light pipe can be selectively inserted into tritium light pipes with different geometric dimensions, and the larger the luminous intensity of the tritium light pipe is, the higher the luminous intensity of the tritium light pipe is, so that the function of adjusting the luminous intensity is realized. The outer diameter of the optical bench of the light source module is consistent with the diameter of a sample tube used by the instrument.
As shown in fig. 4, the light source 2 is an LED light source module, which is composed of a button cell 12, an adjustable constant current source 13, an LED light source 14 and a second collimating lens 15 arranged from top to bottom, and a light outlet 11 is formed at the lower end of the LED light source module. Compared with a tritium light source, the LED light source can provide higher light intensity, can meet the verification and calibration requirements of a luminous immunoassay analyzer with poor sensitivity, can omit an interference filter and a light attenuation sheet, has lower cost, and is a beneficial supplement of the tritium light source. The outer diameter of the optical bench of the light source module is consistent with the diameter of a sample tube used by the instrument.
The spectral neutral filter 5 is a spectral neutral filter in the visible light range, and has transmittance of 50%, 30%, 10%, 3%, 1%, and the like, respectively. When the spectrum neutral filter is used, the spectrum neutral filter is placed between the light source module and the lateral light guide. The diameter of the spectrum neutral filter is larger than the diameter of the light output hole of the light source module.
The lateral light guide 3 outputs the monochromatic light vertically incident from the top to the bottom and the lateral direction at the same time, so that the purpose that one standard device can simultaneously meet the verification and calibration of the luminous immunoassay analyzer in the bottom and lateral detection modes is achieved. The lateral light guide has two implementation modes, one is to use a commercial lateral light guide, and the diameter of the lateral light guide is consistent with the diameter of a sample tube of the instrument, so that the aim of outputting light in multiple directions is fulfilled. The second is to inject 1% -10% protein solution into glass or quartz tube by using the tyndall phenomenon, the diameter of tube is identical with that of the instrument sample tube, and the tube is used as lateral light guide, when light is vertically incident, light output is realized in different directions due to the scattering effect of macromolecule. The length of the lateral light guide or the height of the protein solution in the tube can be adjusted and the length or height should be higher than the optical detection window for lateral detection of the instrument.
The measuring standard can be used for calibrating or evaluating the optical detection indication error, repeatability and linear measuring characteristic index of the instrument. Before evaluation, a lateral light guide with a proper length is selected to cover a detection window; then, the luminous intensity is changed by selecting a proper tritium light source module or adjusting a constant current source of an LED, so that the luminous value I 0 displayed by the instrument reaches about 2/3 of the maximum indication value when the spectrum neutral filter 5 is not inserted into the filter cavity 21. Spectral neutral filters 5 of different transmittances (e.g., T s =50%, 30%, 10%, 3%, 1%, etc.) are then placed in the filter chambers 21, respectively, and the luminous intensity I is measured using a luminescence immunoassay, the transmittance is calculated according to the following formula (1), and compared with a standard value T s of the transmittance of the spectral neutral filter 5, the indication error is calculated as a representation of the instrument indication error.
ΔT＝T-TS (2)
In the case of performing the instrument reproducibility evaluation, the same operation as in the value error calibration evaluation was performed, and the luminescence intensities when the spectral neutral filter 5 was not inserted and when the 1% spectral neutral filter 5 was inserted were measured by using a luminescence immunoassay instrument, and the relative standard deviation of the measurement results was calculated 20 times for each state was repeatedly measured as an indication of the instrument reproducibility.
In the case of performing the linear evaluation of the instrument, the operation is the same as the error calibration evaluation of the indication value, so that the emission value I 0 displayed by the instrument reaches about 2/3 of the maximum indication value when the spectral neutral filter 5 is not inserted into the filter chamber 21. Then, spectral neutral filters (such as 50%, 30%, 10%, 3%, 1% and the like) with different transmittances are respectively placed in the spectral cavity, the luminous intensity I is measured by using a luminous immunoassay analyzer, the actual transmittance of each filter is calculated according to a formula (1), and then linear fitting is carried out on the actual transmittance of each filter to obtain a linear correlation coefficient r serving as the linear representation of the instrument.
The invention and its embodiments have been described above by way of illustration and not limitation, and the invention is illustrated in the accompanying drawings and described in the drawings in which the actual structure is not limited thereto. Therefore, those skilled in the art, having benefit of this disclosure, will appreciate that other embodiments can be devised which do not depart from the spirit and scope of the invention as disclosed herein.
Claims (7)
1. A measurement standard for calibration of a luminescence immunoassay analyzer for bottom or lateral detection, comprising: support (1), light source (2), side direction light guide (3), first interference filter (18), attenuation piece (17) and spectral neutral filter (5), its characterized in that: the light source device comprises a support (1), a light source (2), a light source cavity (21), three pairs of jacks (4), a first interference filter (18), an attenuation sheet (17) and a spectrum neutral filter (5), wherein the upper end of the support (1) is provided with a groove (16), the light source (2) is fixed in the groove (16), the lower end of the support (1) is fixedly provided with a cylindrical lateral light guide (3), the cylindrical barrel between the light source (2) and the lateral light guide (3) is a light filter cavity (21), three pairs of jacks (4) are arranged along the length direction of the light filter cavity (21), each pair of jacks (4) are arranged at symmetrical positions of the circumference of the cylindrical barrel, the first interference filter (18), the attenuation sheet (17) and the spectrum neutral filter (5) are sequentially inserted into the three pairs of jacks (4) from top to bottom along the length direction of the light filter cavity (21), the light outlet holes (11) of the light source (2) face one or more than one or more of the first interference filter (18), the attenuation sheet (17) and the spectrum neutral filter (5).
2. A measurement standard for luminescence immunoassay calibration of bottom or lateral detection of claim 1, wherein: the lateral light guide (3) is a commercial lateral light guide fiber tube or a glass tube or a quartz tube filled with 1% -10% protein solution.
3. A measurement standard for luminescence immunoassay calibration of bottom or lateral detection of claim 2, wherein: the transmittance of the spectral neutral filter (5) is 50%, 30%, 10%, 3% or 1%.
4. A measurement standard for luminescence immunoassay analyzer calibration for bottom or lateral detection of claim 3, wherein: the lateral light guide (3) is fixed at the lower end of the bracket (1) by means of a number of fixing screws (6) along the circumference of the bracket (1).
5. The measurement standard for bottom or lateral detection luminescence immunoassay meter calibration of claim 4, wherein: the lateral light guide (3) is fixed at the lower end of the bracket (1) through threads.
6. A measurement standard for luminescence immunoassay meter calibration for bottom or lateral detection of claim 4 or 5, wherein: the light source (2) is a tritium light source module, the tritium light source module is composed of a tritium light pipe (7), a second interference filter (8), a first collimating lens (9) and a light attenuation sheet (10) which are arranged from top to bottom, and a light outlet hole (11) is formed in the lower end of the tritium light source module.
7. A measurement standard for luminescence immunoassay meter calibration for bottom or lateral detection of claim 4 or 5, wherein: the light source (2) is an LED light source module, the LED light source module is composed of a button battery (12), an adjustable constant current source (13), an LED light source (14) and a second collimating lens (15) which are arranged from top to bottom, and a light outlet hole (11) is formed in the lower end of the LED light source module.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010635924.4A CN111650170B (en) | 2020-07-03 | Metering standard for calibrating luminous immunity analyzer for bottom or side direction detection |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010635924.4A CN111650170B (en) | 2020-07-03 | Metering standard for calibrating luminous immunity analyzer for bottom or side direction detection |
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| Publication Number | Publication Date |
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| CN111650170A CN111650170A (en) | 2020-09-11 |
| CN111650170B true CN111650170B (en) | 2024-07-02 |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN212845025U (en) * | 2020-07-03 | 2021-03-30 | 中国计量科学研究院 | Measurement standard for calibration of luminescence immunoassay analyzer for bottom or lateral detection |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN212845025U (en) * | 2020-07-03 | 2021-03-30 | 中国计量科学研究院 | Measurement standard for calibration of luminescence immunoassay analyzer for bottom or lateral detection |
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