WO2021134302A1 - 用于检测hcv的免疫分析仪、方法及试剂盒 - Google Patents

用于检测hcv的免疫分析仪、方法及试剂盒 Download PDF

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Publication number
WO2021134302A1
WO2021134302A1 PCT/CN2019/130124 CN2019130124W WO2021134302A1 WO 2021134302 A1 WO2021134302 A1 WO 2021134302A1 CN 2019130124 W CN2019130124 W CN 2019130124W WO 2021134302 A1 WO2021134302 A1 WO 2021134302A1
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reagent
solid phase
hcv antigen
component
sample
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PCT/CN2019/130124
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English (en)
French (fr)
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于丽娜
李可
何健文
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深圳迈瑞生物医疗电子股份有限公司
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Priority to CN201980102015.XA priority Critical patent/CN114641689A/zh
Priority to PCT/CN2019/130124 priority patent/WO2021134302A1/zh
Publication of WO2021134302A1 publication Critical patent/WO2021134302A1/zh

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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
    • G01N21/76—Chemiluminescence; Bioluminescence
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N35/02—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor using a plurality of sample containers moved by a conveyor system past one or more treatment or analysis stations

Definitions

  • the present invention relates to the field of immunoassays, in particular to the detection of HCV.
  • HCV Hepatitis C Virus
  • HCV infection will involve the detection of its antibodies and/or antigens.
  • the detection of anti-HCV antibodies often indicates that the patient has a history of past infection, but it cannot distinguish whether the patient is an infected person or a person who has recovered from a previous infection.
  • the detection of HCV antigen can prompt the patient's infection status, and can monitor the patient's treatment process, which is a useful supplementary tool for diagnosis.
  • the detection of HCV antigen can effectively shorten the window period and avoid early infection of the disease.
  • HCV detection kits for specific test items have been developed.
  • a kit on an immunoassay analyzer to test the sample to be tested, only the specific items targeted by the kit can be used.
  • the present invention provides an immunoassay instrument that can selectively detect HCV antigen and/or anti-HCV antigen antibody in a sample to be tested, including:
  • the sample device has a sample storage component and a sample dispensing component, the sample storage component is used to store the sample to be tested, and the sample dispensing component is used to suck the sample to be tested and discharge it into the reaction cup to be added;
  • the reagent device has a reagent storage component and a reagent dispensing component.
  • the reagent storage component is used to store a reagent kit.
  • the reagent kit includes a solid phase reagent and a labeled reagent.
  • the solid phase reagent includes a first solid phase component and a first solid phase component.
  • the labeling reagent includes a first labeling component and a second labeling component, the first labeling component includes a labeled anti-HCV antigen antibody, and the second labeling component includes a labeled anti-human antibody,
  • the reagent dispensing component is used to suck the solid phase components and the labeled components in the reagent box stored on the reagent storage component and discharge them into the reaction cup to which the reagent is to be added;
  • the luminescent substrate dispensing device is connected to the container storing the luminescent substrate and is used to inject the luminescent substrate into the reaction cup where the luminescent substrate is to be added;
  • the reaction device has a plurality of placement positions for placing the reaction cup and is used for incubating the reaction solution in the reaction cup;
  • the optical measurement component is used to perform optical measurement on the reaction solution after the incubation to obtain the detection result of the sample to be tested;
  • the control device is electrically connected to the sample device, the reagent device, the luminescent substrate dispensing device and the light measuring component, and is configured to:
  • test instruction including the type of the substance to be tested
  • Control the reagent dispensing component to add at least one solid phase component corresponding to the type of the substance to be tested into the reaction cup on the reaction device, so that the sample to be tested and the at least one The solid phase components are mixed in the reaction cup and incubated for a period of time, so that the coating body coated on the solid phase carrier can bind to the test substance in the test sample;
  • the reagent dispensing component is controlled to further add at least one marking component corresponding to the type of the substance to be tested into the reaction cup, so that the added marking component is mixed with the mixture in the reaction cup and incubated For a period of time, the labeling body in the added labeling component can be combined with the test substance bound to the coating body coated on the solid carrier to form a complex of the coating body-test substance-labeling body ;
  • the detection result is obtained according to the ratio of the luminescence value measured in the light measuring component to the luminescence threshold value.
  • the immunoassay analyzer is based on a kit capable of responding to instructions containing the types of test substances to select and detect different test substances.
  • the reagent dispensing component first adds only the first solid phase component or at least the first solid phase component to the reaction cup, and the solid phase reagent
  • the anti-HCV antigen antibody coated on the solid phase carrier can specifically bind to the HCV antigen in the sample to be tested; next, the immune analyzer responds to the instruction to further control the reagent dispensing component to select at least the first labeling component or Only the first labeling component is added to the reaction cup, and the detection of HCV antigen is realized by forming a complex of anti-HCV antigen-HCV antigen-anti-HCV antigen antibody.
  • the reagent dispensing part when selecting antibodies against HCV antigens (ie, the test substance is an antibody against HCV antigens) for detection, the reagent dispensing part first adds only the second solid phase component or at least the second solid phase component to the reaction In the cup, the HCV antigen in the solid-phase reagent coated on the solid-phase carrier can specifically bind to the anti-HCV antigen antibody in the sample to be tested; next, the immune analyzer responds to the instruction to further control the reagent dispensing components to select at least The second labeling component or only the second labeling component is added to the reaction cup to form a complex of HCV antigen-anti-HCV antigen antibody-anti-human antibody to realize the detection of antibodies against HCV antigen.
  • the reagent dispensing component first divides the first solid phase component and the second solid phase component
  • the anti-HCV antigen antibody coated on the solid-phase carrier in the solid phase reagent can specifically bind to the HCV antigen in the sample to be tested, and the HCV antigen coated on the solid-phase carrier can interact with The antibody against the HCV antigen in the sample to be tested specifically binds; next, the immune analyzer responds to the instruction to further control the reagent dispensing part to select the first labeling component and the second labeling component to be added to the reaction cup, by forming The complex of antibody-HCV antigen-anti-HCV antigen antibody, HCV antigen-anti-HCV antigen antibody-anti-human antibody, realizes the mixed detection of HCV antigen and anti-HCV antigen antibody.
  • the immunoassay analyzer of the first aspect of the present invention no matter the individual needs to detect HCV antigen, detect antibodies against HCV antigen, mixed detection of HCV antigen and antibodies against HCV antigen, or a combination of the above detection items, it can be passed according to instructions Control the reagent dispensing component to add the corresponding reagent components so as to be realized flexibly and conveniently.
  • the reagent storage part of the immune analyzer does not need to store additional kits, so testers do not need to be equipped with multiple kits, and will not waste kits due to different usage frequencies, thereby reducing the cost of testing.
  • the anti-HCV antigen antibody and the HCV antigen coated on the solid phase carrier in the solid phase reagent are present in separate packages; in the labeled reagent, the labeled anti-HCV antigen antibody
  • the anti-human antibody and anti-human antibody exist in separate aliquots or pre-mixed forms.
  • the anti-HCV antigen antibody and the HCV antigen coated on the solid phase carrier in the solid phase reagent are present in a pre-mixed form; in the labeling reagent, the labeled anti-HCV antigen antibody and Anti-human antibodies exist in separate aliquots.
  • the present invention provides an immunoassay method for detecting HCV, which includes the following steps:
  • the sample to be tested is mixed with the anti-HCV antigen antibody and HCV antigen coated on the solid phase carrier and incubated for a period of time, so that the anti-HCV antigen antibody and HCV antigen coated on the solid phase carrier can interact with the test specimen. Binding of the substance to be tested in the sample;
  • the anti-HCV antigen antibody on the solid phase carrier binds to the test substance bound to the HCV antigen to form a complex
  • a luminescent substrate is added to the washed complex to detect the mixed detection value of the anti-HCV antigen antibody and the HCV antigen in the sample to be tested.
  • the solid phase reagent is mixed with the sample to be tested and incubated, and the solid phase reagent includes an anti-HCV antigen antibody and an HCV antigen coated on a solid phase carrier, which are respectively It can specifically bind to alternative test substances, namely HCV antigens and antibodies against HCV antigens.
  • the labeled component is mixed with the mixture of the sample and the solid phase reagent and incubated to mix the detection HCV antigen and the antibody against the HCV antigen.
  • the method of the present invention can finally obtain a mixed detection result, which can perform mixed detection of HCV antigen and anti-HCV antigen antibody in the sample, and improve the detection efficiency.
  • the anti-HCV antigen antibody and the HCV antigen coated on the solid carrier are added to the test sample, mixed and incubated.
  • the anti-HCV antigen antibody and the HCV antigen coated on the solid carrier are added to the test sample in a pre-mixed form, mixed and incubated.
  • labeled anti-HCV antigen antibodies and anti-human antibodies are added to the washed mixture, mixed and incubated.
  • the labeled anti-HCV antigen antibody and the anti-human antibody are added to the washed mixture in a pre-mixed form, mixed and incubated.
  • the present invention provides an immunoassay method for selective detection of HCV, which includes the following steps:
  • At least one of the first solid phase component and the second solid phase component is selected from the solid phase reagent, and the sample to be tested is mixed with the selected solid phase component and incubated for a period of time, so that At least one coating body coated on the solid-phase carrier can bind to the test substance in the test sample;
  • At least one of the first labeling component and the second labeling component is selected from the labeling reagent, and the selected labeling component is added to the washed mixture and incubated so that the added labeling group
  • the labeling body in the component can be combined with the test substance bound to at least one coating body coated on the solid carrier to form a complex
  • the first solid phase component is an antibody against HCV antigen coated on a solid phase carrier
  • the second solid phase component is an HCV antigen coated on a solid phase carrier
  • the first label The component is a labeled anti-HCV antigen antibody
  • the second labeled component is a labeled anti-human antibody.
  • the first solid phase component and/or the second solid phase component are selected from the solid phase reagents according to the requirements of the type of substance to be tested, and the selected The solid phase component is mixed with the sample to be tested and incubated, so that at least one coating body coated on the solid phase carrier can bind to the substance to be tested.
  • the first labeling component and/or the second labeling component from the labeling reagent, and mix and incubate the selected labeling component with the mixture of the sample and the solid phase reagent ,
  • selective detection of HCV antigen detection of anti-HCV antigen antibodies, mixed detection of HCV antigen and anti-HCV antigen antibodies.
  • the present invention provides a kit for detecting HCV, including:
  • Solid-phase reagents including anti-HCV antigen antibodies and HCV antigens coated on a solid-phase carrier;
  • Labeling reagents including labeled anti-HCV antigen antibodies and anti-human antibodies.
  • the anti-HCV antigen antibody and the HCV antigen coated on the solid-phase carrier in the solid-phase reagent are in the form of separate packages.
  • the anti-HCV antigen antibody and the HCV antigen coated on the solid phase carrier in the solid phase reagent are present in a pre-mixed form.
  • the labeled anti-HCV antigen antibody and the anti-human antibody in the labeling reagent are present in separate aliquots.
  • the labeled anti-HCV antigen antibody and the anti-human antibody in the labeling reagent are present in a pre-mixed form.
  • the kit provided in the fourth aspect of the present invention can be applied to the immune analyzer provided in the first aspect of the present invention and the immunoassay methods provided in the second and third aspects of the present invention.
  • the anti-human antibody is selected from at least one of an anti-human IgG antibody and an anti-human IgM antibody.
  • the concentration of each substance in the solid phase reagent and the labeling reagent is designed such that each of the solid phase reagents is used in the same reaction system.
  • a coating body coated on a solid-phase carrier and a corresponding labeling body in the labeling reagent are separately tested for internal reference products (for example, internal reference products defined according to the ISO18113-1:2009 standard),
  • the corresponding luminescence thresholds are basically the same.
  • Figure 1 shows a schematic diagram of an immunoassay system according to an embodiment of the present invention
  • Fig. 2 shows a schematic structural diagram of a control device according to an embodiment of the present invention.
  • the present invention designs a method for selectively detecting antigens and/or antibodies, that is, based on the same kit, it can selectively realize antigen detection, antibody detection, or mixed detection of antigen and antibody. Users can select one or several of them for testing according to their needs.
  • the term "coating body” refers to a substance coated on a solid-phase carrier, which can bind to the substance to be tested in the sample to be tested.
  • labeled substance refers to a substance labeled with a marker, which can bind to the substance to be tested in the sample to be tested.
  • solid support refers to a solid surface to which antigens or antibodies can be attached.
  • solid-phase carrier used in the present invention
  • commercial solid-phase carriers and any solid-phase carrier that can be used in immunoassays can be used in the present invention.
  • Exemplary solid phase carriers can be magnetic beads (such as carboxyl magnetic beads), enzyme-labeled plates, plastic plates, plastic tubes, latex beads, agarose beads, glass, nitrocellulose membranes, nylon membranes, silica plates, or micro Chip, but the present invention is not limited to this.
  • the anti-HCV antigen antibody and HCV antigen coated on the solid phase carrier can be produced in the following manner: On the one hand, the HCV antigen antibody and HCV antigen can be separately coated on different solid phase carriers Then, the coated solid phase carrier is mixed. On the other hand, it is also possible to mix the HCV antigen antibody and HCV antigen first, and then coat it with the solid phase carrier.
  • markers that can be used in the embodiments of the present invention are well known to those skilled in the art, such as alkaline phosphatase (ALP), peroxidase, microperoxidase, horseradish peroxidase, ⁇ -half Enzymes such as lactosidase, glucose oxidase and glucose 6-phosphate dehydrogenase; fluorescein isothiocyanate, tetramethylrhodamine isothiocyanate, fluorescein, rhodamine, europium and green fluorescent protein, etc.
  • ALP alkaline phosphatase
  • peroxidase microperoxidase
  • horseradish peroxidase ⁇ -half Enzymes
  • lactosidase glucose oxidase and glucose 6-phosphate dehydrogenase
  • fluorescein isothiocyanate tetramethylrhodamine isothiocyanate
  • fluorescein, rhodamine europium
  • Fluorescent substances such as luminol, isoluminol, phenanthridinium and acridinium esters; coenzymes such as NAD; biotin; radioactive substances such as 35 S, 14 C, 32 P, 131 I and 125 I , but the present invention is not limited to this.
  • a suitable luminescent substrate can be selected according to the type of label used to generate a detectable signal.
  • alkaline phosphatase 3-(2-helicaladamantane)-4-methoxy-4-(3-phosphooxy)-phenyl-1,2-dioxide can be used.
  • Cycloethane is used as a luminescent substrate.
  • the substrate will be decomposed by alkaline phosphatase to remove a phosphate group and generate an unstable intermediate product.
  • the intermediate product generates methyl meta-oxybenzoate anion through intramolecular electron transfer.
  • chemiluminescence is generated.
  • the number of photons generated in the reaction is measured by a photomultiplier tube, and the amount of photons generated is proportional to the content of the detected substance in the sample.
  • the embodiments of the present invention are applicable to various methods such as ELISA, chemiluminescence, electrochemiluminescence, POCT, immunochromatography, up-conversion luminescence, and down-conversion luminescence.
  • ROC receiver operating characteristic
  • ROC curve refers to the curve obtained by dividing the diagnostic test result into several critical points, with the sensitivity corresponding to each critical point as the ordinate and the specificity as the abscissa.
  • ROC curve is an effective tool for comprehensive and accurate evaluation of diagnostic tests.
  • Another function of the ROC curve is to determine the optimal threshold for detection.
  • ROC curve method to determine the critical point In most cases, select the point on the curve as close to the upper left as possible to determine the critical point as the best.
  • the antigen of the embodiment of the present invention may exist in the form of, for example, a multimer, a recombinant antigen, a natural antigen, an antigen fragment, or an antigen peptide.
  • the antibodies of the embodiments of the present invention may exist in the form of monoclonal antibodies, polyclonal antibodies, recombinant antibodies, chimeric antibodies, humanized antibodies, and antigen-binding fragments of antibodies, for example.
  • the antibodies may be derived from mice, rabbits, goats, sheep, and chickens, but the present invention is not limited thereto.
  • anti-human antibody refers to an antibody that can specifically bind to a human antibody.
  • the labeling substance in the labeling reagent may be connected to the labeling substance by a method conventional in the art (for example, chemical bonding).
  • the ratio of the detection value to the luminescence threshold is used to determine whether the result is positive or negative. For example, when the ratio is greater than or equal to 1.1, the judgment result is positive, indicating that the test result of at least one substance to be tested is positive. When the ratio is between 0.9 and 1.1, the judgment result is a gray zone, which is neither positive or negative; when the ratio is less than 0.9, the judgment result is negative, indicating that the test results for all the tested substances are negative.
  • the concentration of each component in the solid phase reagent and the labeling reagent is designed such that each of the solid phase reagents is used to coat the solid phase in the same reaction system.
  • the corresponding luminescence thresholds are basically the same.
  • the solid phase component and the labeling component corresponding to a certain substance to be tested are used to separately detect the internal reference defined in ISO 18113-1:2009, and the concentration of the labeling component is maintained as A constant value is used to adjust the concentration of the solid phase component to adjust the luminescence threshold to a preset value; or, to maintain the concentration of the solid phase component at a constant value, to adjust the concentration of the marker component to adjust the luminescence threshold to the preset value.
  • internal reference refers to the standard and basis determined by the system composition, which has the definition as in the international standard ISO 18113-1:2009 and can be obtained according to the standard.
  • Internal reference materials are samples used by medical device manufacturers to verify product performance. They are the most important criteria and basis for product selection, preparation, identification, and determination of raw material quality standards, product production process determination, reaction system composition, reaction conditions, etc. .
  • the identification sample of the company's internal reference product is a sample that judges the value or quantity of a specific disease, state, or the boundary between the existence and non-existence of the measurement.
  • substantially the same means that the relative deviation is within ⁇ 10%, such as within ⁇ 5%, within ⁇ 3%, within ⁇ 2%, or within ⁇ 1%.
  • the embodiment of the present invention provides an immunoassay analyzer that can selectively detect HCV antigens and/or antibodies against HCV antigens in a sample to be tested.
  • the immunoassay analyzer includes a sample device 10, a reagent device 20, a reaction device 30, a light measuring component 40, and a control device 50.
  • the immune analyzer may also include a display part (not shown).
  • the sample device 10 is used to carry the sample to be tested, aspirate the sample and provide it to the reaction device 30.
  • the sample device 10 includes a sample storage part 11 and a sample dispensing part 12.
  • the sample storage component 11 is used to store samples to be tested.
  • the sample storage component 11 may include a sample distribution module (SDM, Sample Delivery Module) and a front-end track.
  • the sample storage component 11 may also be a sample tray.
  • the sample tray includes a plurality of sample positions such as sample tubes. By rotating the tray structure of the sample tray, the sample can be scheduled to the corresponding position, for example, for The position where the sample dispensing part 12 sucks the sample.
  • the sample dispensing component 12 is used to aspirate the sample and discharge it into the reaction cup to be added.
  • the sample dispensing component 12 may include, for example, a sample needle.
  • the sample needle performs a two-dimensional or three-dimensional movement in space through a two-dimensional or three-dimensional drive mechanism, so that the sample needle can move to aspirate the sample carried by the sample storage component 11, and Move to the reaction cup to be added, and discharge the sample into the reaction cup.
  • the reagent device 20 is used to carry reagents, and the reagents are sucked and supplied to the reaction device 30.
  • the reagent device 20 includes a reagent storage part 13 and a reagent dispensing part 14.
  • the reagent storage part 13 is used to store a reagent cartridge.
  • the reagent storage component 13 may be a reagent tray.
  • the reagent tray is arranged in a disc-shaped structure and has multiple positions for carrying reagent containers.
  • the reagent storage component 13 can rotate and drive the reagent container it carries to rotate. It is used to rotate the reagent container to a specific position, for example, a position where the reagent is sucked by the reagent dispensing part 14.
  • the number of reagent storage parts 13 may be one or more.
  • the reagent dispensing part 14 is used for sucking the reagents in the reagent box and discharging them into the reaction cup to be added with the reagents.
  • the reagent dispensing part 14 may include a reagent needle, and the reagent needle can move in a two-dimensional or three-dimensional space through a two-dimensional or three-dimensional drive mechanism, so that the reagent needle can move to absorb the reagent storage part 13 The loaded reagent, and move to the reaction cup where the reagent is to be added, and discharge the reagent into the reaction cup.
  • the reagent storage component 13 is used to store reagent kits.
  • the kit includes a solid phase reagent and a labeling reagent.
  • the solid phase reagent includes a first solid phase component and a second solid phase component, the first solid phase component includes an anti-HCV antigen antibody coated on a solid phase carrier, and the second solid phase component includes HCV antigen coated on a solid carrier.
  • the labeling reagent includes a first labeling component and a second labeling component.
  • the first labeling component includes a labeled anti-HCV antigen antibody, and the second labeling component includes a labeled anti-human antibody.
  • the reaction device 30 has at least one placement position for placing the reaction cup and incubating the reaction solution in the reaction cup.
  • the reaction device 30 may be a reaction disc, which is arranged in a disc-shaped structure and has one or more placement positions for placing reaction cups. The reaction disc can rotate and drive the reaction cup in its placement position to rotate for The reaction cup is arranged in the reaction tray and the reaction solution in the incubation reaction cup is incubated.
  • the optical measurement component 40 is used to perform optical measurement on the reaction solution after the incubation to obtain reaction data of the sample.
  • the light measuring component 40 detects the luminous intensity of the reaction solution to be measured, and calculates the concentration of the component to be measured in the sample through a calibration curve.
  • the optical sensing component 40 is separately arranged outside the reaction device 30.
  • the immunoassay analyzer also includes a luminescent substrate dispensing device (not shown).
  • the luminescent substrate dispensing device is connected with the container storing the luminescent substrate and is used for injecting the luminescent substrate into the reaction cup to be added with the luminescent substrate.
  • the control device 50 includes at least: a processing component 51, a RAM 52, a ROM 53, a communication interface 54, a memory 56 and an I/O interface 55.
  • the processing component 51, RAM 52, ROM 53, communication interface 54, memory 56 and the I/O interface 55 communicate through the bus 57.
  • the processing component may be a CPU, GPU, or other chips with computing capabilities.
  • the memory 56 is loaded with various computer programs such as an operating system and application programs for the processor component 51 to execute, and data required to execute the computer programs. In addition, during the sample detection process, any data that needs to be stored locally can be stored in the memory 56.
  • the I/O interface 55 is composed of a serial interface such as USB, IEEE1394 or RS-232C, a parallel interface such as SCSI, IDE or IEEE1284, and an analog signal interface composed of a D/A converter and an A/D converter.
  • An input device composed of a keyboard, a mouse, a touch screen or other control buttons is connected to the I/O interface 55, and the user can use the input device to directly input data to the control device 50.
  • the I/O interface 55 can also be connected to a display with display function, such as: LCD screen, touch screen, LED display screen, etc., and the control device 50 can output the processed data as image display data to the display for display, for example : Analyze data, instrument operating parameters, etc.
  • the communication interface 54 is an interface that can be any currently known communication protocol.
  • the communication interface 54 communicates with the outside world through the network.
  • the control device 50 can transmit data to any device connected through the network through the communication interface 54 in a certain communication protocol.
  • the control device 50 is configured to receive a test instruction, the test instruction includes the type of the substance to be tested, and the following steps are executed in response to the test instruction:
  • the control reagent dispensing component 14 adds at least one solid phase component corresponding to the type of the substance to be tested in the reagent box into the reaction cup on the reaction device 30, so that the sample to be tested and the added At least one solid phase component is mixed in the reaction cup and incubated for a period of time, so that the coating body on the solid phase carrier can bind to the test substance in the test sample;
  • the control reagent dispensing component 14 further adds the labeled component corresponding to the type of the substance to be tested in the reagent box into the reaction cup, so that the added labeled component is mixed with the mixture in the reaction cup and incubated for a period of time Time so that the labeling body in the added labeling component can bind to the test substance bound on the coating body in the added solid phase reagent;
  • the detection result is obtained according to the ratio of the luminescence value measured in the light measuring component 40 to the luminescence threshold value.
  • the immune analyzer provided by the embodiment of the present invention can selectively detect different substances to be tested based on a kit based on user requirements, thereby improving detection flexibility and meeting user requirements in different scenarios.
  • Anti-HCV antigen antibodies are from Meridian Life Science;
  • Anti-human antibodies come from: Jackson ImmunoResearch;
  • HCV antigen comes from Meridian Life Science
  • Alkaline phosphatase comes from Roche Pharmaceuticals
  • the magnetic beads come from Thermo Fisher.
  • the antigen/antibody is pretreated, and the protective components in the buffer matrix are removed by dialysis.
  • the coating is carried out at a ratio of 1-40 ⁇ g of antigen/antibody (preferably 5-30 ⁇ g) per mg of magnetic beads.
  • the carboxyl group on the surface of the magnetic bead is coupled with the amino group of the antigen/antibody under the catalysis of EDC/NHS.
  • the anti-human antibody and anti-HCV antigen antibody are labeled with signal markers.
  • the signal marker is alkaline phosphatase. Choose 50mM MES pH 6.0 buffer to dilute the anti-human antibody signal markers and anti-HCV antigen antibody signal markers to prepare labeled components.
  • the sample and solid phase reagent are added to the reaction tube, and incubated at 37°C for 10 minutes, so that the antigen/antibody on the solid phase surface can bind to the corresponding test substance in the sample.
  • the substance bound to the solid phase will be placed in a magnetic field and be attracted, the substance bound to the solid phase of the magnetic beads is retained, and the unbound substance is washed and removed.
  • the second step select the required labeled components and add them to the reaction tube to incubate, mix, and incubate at 37°C for 10 minutes to bind to the conjugate formed in the first step. After the incubation in the reaction tube is completed, the complex is attracted by the magnetic field, and other unbound substances are washed and removed.
  • the third step is to add AMPPD to the reaction tube to generate chemiluminescence.
  • the number of photons produced by the reaction is measured by a photomultiplier tube to obtain the chemiluminescence signal value of the sample.
  • the luminescence signal value is divided by the threshold value to obtain the COI value. Compare the COI value of the test result of the sample with the reference value (reference value 1.10). If it is greater than or equal to 1.1, it means that one or more of the test substances in the sample is positive; if it is less than 0.90, it means that the test substances in the sample are all Is negative. The COI is between 0.90 and 1.10, and the result is a gray zone (indeterminate).
  • the negative coincidence rate refers to the ratio of the number of samples judged to be negative using the test method of the embodiment of the present invention to the negative samples actually participating in the evaluation
  • the positive coincidence rate refers to the test using the embodiment of the present invention
  • the method obtains the proportion of the number of positive samples judged to be positive to the positive samples actually participating in the evaluation; the true negative and positive results of the samples come from the diagnosis results of the hospital.
  • the magnetic microspheres coated with HCV antigen obtained by "preparation of solid phase coating” as the solid phase component the anti-human antibody with alkaline phosphatase obtained by "preparation of labeling component”
  • the positive and negative coincidence rates of antibodies against HCV antigens are tested under different luminescence thresholds according to the "detection steps".
  • the magnetic microspheres coated with antibodies against HCV antigens obtained from the "preparation of solid phase coating” are used as the solid phase components, and those obtained from the “preparation of labeled components” are alkaline
  • the phosphatase anti-HCV antigen antibody is used as the labeling component, and the positive coincidence rate and negative coincidence rate of the HCV antigen are tested under different luminescence thresholds according to the "detection step". The results are summarized in Table 1.
  • Preparation of solid phase reagent use the magnetic microspheres coated with HCV antigen and the magnetic microspheres coated with antibodies against HCV antigen obtained from "Preparation of solid phase coating" as solid phase reagents;
  • Preparation of labeling component 1 According to "Preparation of labeling component", use the prepared anti-human antibody with alkaline phosphatase, and match it with the corresponding solid phase component to determine the internal reference. Select the dilution corresponding to the luminescence threshold of 70,000 (allowing ⁇ 3% deviation) as the dilution of labeled component 1;
  • Preparation of labeling component 2 According to "Preparation of labeling component", use the prepared anti-HCV antigen antibody with alkaline phosphatase, and match it with the corresponding solid phase component to determine the internal reference. The dilution corresponding to the luminescence threshold of 70,000 (allowing ⁇ 3% deviation) is selected as the dilution of labeled component 2.
  • a total of 1,000 samples with clear clinical diagnosis results were selected. Among them, 670 were HCV antibody and antigen negative samples, and 330 were HCV antibody or antigen positive samples (318 were HCV antibody positive and 211 were HCV antigen positive).
  • the positive coincidence rate is 100%, the negative coincidence rate is 99.06%, and the overall coincidence rate is 99.77%; when the HCV antigen is tested alone, the positive coincidence rate is 100%, and the negative coincidence rate is 100%. 99.53%, the overall coincidence rate is 99.90%; mixed detection of HCV antigen and antibody, the positive coincidence rate is 100%, the negative coincidence rate is 98.48%, and the overall coincidence rate is 99.95%.
  • the embodiment of the present invention can realize the selective detection of HCV antigen and/or anti-HCV antigen in the sample to be tested.

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Abstract

可选择性检测待测样本中的HCV抗原和/或抗HCV抗原的抗体的免疫分析仪，该免疫分析仪配套一个试剂盒可根据用户需求，实现不同HCV项目的选择性检测。还涉及免疫分析方法及试剂盒。

Description

用于检测HCV的免疫分析仪、方法及试剂盒 技术领域
本发明涉及免疫分析领域，具体涉及HCV的检测。
背景技术
Michael Houghto等于1989年分离并命名了丙型肝炎病毒(Hepatitis C Virus,HCV)，HCV的慢性感染可能导致肝脏慢性炎症坏死和肝纤维化，部分患者可发展为肝硬化甚至肝细胞癌，对患者的健康和生命危害极大，已成为严重的社会和公共卫生问题。据世界卫生组织统计，全球HCV的感染率约为3％，估计约1.7亿人感染了HCV。
在临床应用中，对于HCV的感染，会涉及对其抗体和/或抗原进行检测。其中抗HCV抗体的检测往往提示患者存在既往感染史，但不能够鉴区分患者是感染者或者是既往感染痊愈者。对HCV抗原的检测可提示患者的感染状态，并能监测患者治疗过程，是诊断有用的补充工具。此外对HCV抗原的检测可以有效的缩短窗口期，避免疾病的早期感染。
目前，已开发了针对特定测试项目的HCV的检测试剂盒。然而，在免疫分析仪上使用这样的试剂盒对待测样本进行检测时，仅能够试剂盒所针对的特定项目。由于抗HCV抗体、HCV抗原以及抗体和抗原的混合检测的使用频率并不相同，这就导致了医院需要准备大量不同种类的试剂盒以应对不同需求，同时不可避免地造成了医疗成本的增加。
据此，在使用免疫分析仪进行HCV检测时，存在着按照用户需求，灵活地、便捷地选择检测项目的强烈需求。
发明内容
为了解决当前免疫分析仪在基于同一试剂盒对HCV进行检测的情况下，无法根据需求选择合适的检测项目(即，仅HCV抗原、仅抗HCV抗原的抗体、HCV抗原和抗HCV抗原的抗体混合)的问题。本发明人对免疫分析仪的工作模式及配套的试剂盒进行了研究，从而实现本发明。
在第一方面，本发明提供了一种可选择性检测待测样本中的HCV抗原和/或抗HCV抗原的抗体的免疫分析仪，包括：
样本装置，具有样本存储部件和样本分注部件，所述样本存储部件用于存储待测样本，所述样本分注部件用于吸取所述待测样本并排放到待加样的反应杯中；
试剂装置，具有试剂存储部件和试剂分注部件，所述试剂存储部件用于存储试剂盒，所述试剂盒包括固相试剂和标记试剂，所述固相试剂包括第一固相组分和第二固相组分，所述第一固相组分包括包被在固相载体上的抗HCV抗原的抗体，所述第二固相组分包括包被在固相载体上的HCV抗原，所述标记试剂包括第一标记组分和第二标记组分，所述第一标记组分包括带标记物的抗HCV抗原的抗体，所述第二标记组分包括带标记物的抗人抗体，所述试剂分注部件用于吸取所述试剂存储部件上存储的试剂盒中的固相组分和标记组分并排放到待加试剂的反应杯中；
发光底物分注装置，与存储发光底物的容器连接并用于将发光底物注入到待加发光底物的反应杯中；
反应装置，具有多个用于放置所述反应杯的放置位并用于孵育所述反应杯中的反应液；
光测部件，用于对孵育完成的反应液进行光测定，以得到待测样本的检测结果；
控制装置，与所述样本装置、所述试剂装置、所述发光底物分注装置 和所述光测部件电连接，配置用于：
接收测试指令，所述测试指令包括待测物质的类型；
响应于所述测试指令：
控制所述样本分注部件将所述样本存储部件中的待测样本加入到所述反应装置上的反应杯中；
控制所述试剂分注部件将所述待测物质的类型所对应的至少一种固相组分加入到所述反应装置上的反应杯中，以使所述待测样本与所述至少一种固相组分在所述反应杯中混合并孵育一段时间，使得包被在固相载体上的包被体能与所述待测样本中的所述待测物质结合；
控制所述试剂分注部件进一步将所述待测物质的类型所对应的至少一种标记组分加入所述反应杯，以使所加入的标记组分与所述反应杯中的混合物混合并孵育一段时间，使得所加入的标记组分中的标记体能与包被在固相载体上的包被体上结合的所述待测物质结合，形成包被体-待测物质-标记体的复合物；
控制所述发光底物分注装置将发光底物加入所述反应杯中；以及
根据所述光测部件中所测得发光值与发光阈值的比值获得检测结果。
需要说明的是，根据本发明第一方面的免疫分析仪基于一种试剂盒能够响应包含待测物质类型的指令，选择检测不同的待测物质。例如，当选择对HCV抗原(即待测物质为HCV抗原)进行检测时，试剂分注部件首先将仅第一固相组分或至少第一固相组分加入到反应杯中，固相试剂中包被在固相载体上的抗HCV抗原的抗体能与待测样本中的HCV抗原特异性结合；接下来，免疫分析仪响应指令进一步控制试剂分注部件以选择至少第一标记组分或仅第一标记组分加入到该反应杯中，通过形成抗HCV抗原的抗体-HCV抗原-抗HCV抗原的抗体的复合物，实现对HCV抗原的检测。 又例如，当选择对抗HCV抗原的抗体(即待测物质为抗HCV抗原的抗体)进行检测时，试剂分注部件首先将仅第二固相组分或至少第二固相组分加入到反应杯中，固相试剂中包被在固相载体上的HCV抗原能与待测样本中的抗HCV抗原的抗体特异性结合；接下来，免疫分析仪响应指令进一步控制试剂分注部件以选择至少第二标记组分或仅第二标记组分加入到该反应杯中，通过形成HCV抗原-抗HCV抗原的抗体-抗人抗体的复合物，实现对抗HCV抗原的抗体的检测。再例如，当选择对HCV抗原和抗HCV抗原(即待测物质为HCV抗原和抗HCV抗原的抗体)进行混合检测时，试剂分注部件首先将第一固相组分和第二固相组分加入到反应杯中，固相试剂中包被在固相载体上的抗HCV抗原的抗体能与待测样本中的HCV抗原特异性结合，而包被在固相载体上的HCV抗原能与待测样本中的抗HCV抗原的抗体特异性结合；接下来，免疫分析仪响应指令进一步控制试剂分注部件以选择第一标记组分和第二标记组分加入到该反应杯中，通过形成抗体-HCV抗原-抗HCV抗原的抗体、HCV抗原-抗HCV抗原的抗体-抗人抗体的复合物，实现对HCV抗原和抗HCV抗原的抗体的混合检测。
采用本发明第一方面的免疫分析仪，无论个体需要检测HCV抗原、检测抗HCV抗原的抗体、混合检测HCV抗原和抗HCV抗原的抗体，又或是上述检测项目的组合，都可以通过根据指令控制试剂分注部件加入对应的试剂组分从而灵活地、便捷地实现。同时，该免疫分析仪的试剂存储部件中不需要储存额外的试剂盒，因此测试人员无需配备多种试剂盒，也不会因为使用频率不同而造成试剂盒的浪费，从而降低了检测成本。
在一些实施方式中，固相试剂中包被在固相载体上的抗HCV抗原的抗体和HCV抗原以单独分装的形式存在；在所述标记试剂中，带标记物的抗HCV抗原的抗体和抗人抗体以单独分装或预先混合的形式存在。
在一些实施方式中，固相试剂中包被在固相载体上的抗HCV抗原的抗 体和HCV抗原以预先混合的形式存在；在所述标记试剂中，带标记物的抗HCV抗原的抗体和抗人抗体以单独分装的形式存在。通过使用单一的固相试剂及选择标记试剂中的一种或更多种标记组分，实现了便捷、成本较低的选择性免疫分析。
在第二方面，本发明提供了一种检测HCV的免疫分析方法，包括以下步骤：
将待测样本与包被在固相载体上的抗HCV抗原的抗体和HCV抗原混合并孵育一段时间，使得包被在固相载体上的抗HCV抗原的抗体和HCV抗原能与所述待测样本中的待测物质结合；
对所述待测样本和所述固相试剂的混合物进行清洗，除去未结合的物质；
向经清洗的混合物中加入带标记物的抗HCV抗原的抗体和带标记物的抗人抗体并孵育，使得带标记物的抗HCV抗原的抗体和带标记物的抗人抗体能与包被在固相载体上的抗HCV抗原的抗体和HCV抗原上结合的所述待测物质结合，形成复合物；
对所述复合物进行清洗，除去未结合的物质；
在经清洗的复合物中加入发光底物，以检测所述待测样本中抗HCV抗原的抗体和HCV抗原的混合检测值。
需要说明的是，在本发明第二方面的方法中，固相试剂与待测样本进行混合并孵育，固相试剂包含包被在固相载体上的抗HCV抗原的抗体和HCV抗原，它们分别能与备选的待测物，即HCV抗原、抗HCV抗原的抗体特异性结合。接下来，将标记组分与样本和固相试剂的混合物进行混合并孵育，以混合检测HCV抗原和抗HCV抗原的抗体。换句话说，采用本发明的方法能够最终得到一个混合检测结果，能够对样本中的HCV抗原、抗HCV抗原的抗体进行混合检测，提高检测效率。
在一些实施方式中，包被在固相载体上的抗HCV抗原的抗体和HCV抗原被分别加入到所述待测样本中进行混合并孵育。
在一些实施方式中，包被在固相载体上的抗HCV抗原的抗体和HCV抗原以预先混合的形式被加入到所述待测样本中进行混合并孵育。
在一些实施方式中，带标记物的抗HCV抗原的抗体和抗人抗体被分别加入到所述经清洗的混合物中进行混合并孵育。
在一些实施方式中，带标记物的抗HCV抗原的抗体和抗人抗体以预先混合的形式被加入到所述经清洗的混合物中进行混合并孵育。
在第三方面，本发明提供了一种选择性检测HCV的免疫分析方法，包括以下步骤：
根据待测物质的类型，从固相试剂中选择第一固相组分和第二固相组分中至少一种，将待测样本与所选择的固相组分混合并孵育一段时间，使得包被在固相载体上的至少一种包被体能与所述待测样本中的所述待测物质结合；
对所述待测样本和所选择的固相组分的混合物进行清洗，除去未结合的物质；
根据待测物质的类型，从标记试剂中选择第一标记组分和第二标记组分中至少一种，向经清洗的混合物中加入所选择的标记组分并孵育，使得所加入的标记组分中的标记体能与包被在固相载体上的至少一种包被体上结合的所述待测物质结合，形成复合物；
对所述复合物进行清洗，除去未结合的物质；
在经清洗的复合物中加入发光底物，以检测所述待测样本中所述待测物质的检测值，
其中，所述第一固相组分为包被在固相载体上的抗HCV抗原的抗体，所述第二固相组分为包被在固相载体上的HCV抗原，所述第一标记组分为 带标记物的抗HCV抗原的抗体，所述第二标记组分为带标记物的抗人抗体。
需要说明的是，在本发明第三方面的方法中，根据待测物质的类型的需求，从固相试剂中选择第一固相组分和/或第二固相组分，将所选择的固相组分与待测样本进行混合并孵育，使得包被在固相载体上的至少一种包被体能与待测物质结合。接下来，根据待测物质的类型的需求，从标记试剂中选择第一标记组分和/或第二标记组分，将所选的标记组分与样本和固相试剂的混合物进行混合并孵育，以选择性检测HCV抗原、检测抗HCV抗原的抗体、混合检测HCV抗原和抗HCV抗原的抗体。
在第四方面，本发明提供了一种检测HCV的试剂盒，包括：
固相试剂，包含包被在固相载体上的抗HCV抗原的抗体和HCV抗原；
标记试剂，包含带标记物的抗HCV抗原的抗体和抗人抗体。
在一些实施方式中，固相试剂中包被在固相载体上的抗HCV抗原的抗体和HCV抗原以单独分装的形式存在。
在一些实施方式中，固相试剂中包被在固相载体上的抗HCV抗原的抗体和HCV抗原以预先混合的形式存在。
在一些实施方式中，标记试剂中带标记物的抗HCV抗原的抗体和抗人抗体以单独分装的形式存在。
在一些实施方式中，标记试剂中带标记物的抗HCV抗原的抗体和抗人抗体以预先混合的形式存在。
本发明第四方面提供的试剂盒可应用于本发明第一方面提供的免疫分析仪以及本发明第二方面和第三方面提供免疫分析方法。
在本发明第一方面至第四方面中，具体地，抗人抗体选自抗人IgG抗体和抗人IgM抗体中的至少一种。
在本发明第一方面至第四方面中，具体地，所述固相试剂和所述标记试剂中各物质的浓度被设计为使得，在同一个反应体系下采用所述固相试 剂中的每一种包被在固相载体上的包被体和所述标记试剂中对应带标记物的标记体单独对内部参考品(例如根据ISO18113-1:2009标准定义的内部参考品)进行检测时，所对应的发光阈值基本上相同。
附图说明
图1示出了本发明实施方式的免疫分析系统的示意图；
图2示出了本发明实施方式的控制装置的结构示意图。
具体实施方式
下文将结合具体实施方式和实施例，具体阐述本发明，本发明的优点和各种效果将由此更加清楚地呈现。本领域技术人员应理解，这些具体实施方式和实施例是用于说明本发明，而非限制本发明。
在整个说明书中，除非另有特别说明，本文使用的术语应理解为如本领域中通常所使用的含义。因此，除非另有定义，本文使用的所有技术和科学术语具有与本发明所属领域技术人员的一般理解相同的含义。若存在矛盾，本说明书优先。
如前文所述，在进行HCV检测时，抗体、抗原以及抗体和抗原的混合检测各自有其应用范围，有的检测项目用于筛查，而有的测试仅用于特殊场景。同时，这些项目在临床有着不同的使用频率。
上述现状给测试方，尤其是一些就诊人数较少的医院，带来了极大不便。一些测试量较少的项目在试剂盒有效期末仍旧有大量的测试数未使用，导致医疗成本的增加。另一方面，测试方需要同时购置多种试剂盒及其配套的校准品和质量控制品，这也是非常不方便的。
对此，本发明设计了一种可选择性检测抗原和/或抗体的方法，即基于同一个试剂盒，可以选择性的实现抗原检测、抗体检测，或抗原和抗体的混合检测。用户可以根据自己的需求，选择其中的某一项或几项进行检测。
在本发明实施方式中，术语“包被体”是指固相载体上所包被的物质，其能与待测样本中的待测物质结合。
在本发明实施方式中，术语“标记体”是指标记有标记物的物质，其能与待测样本中的待测物质结合。
在本发明实施方式中，术语“固相载体”、“固相支持物”、“固体支持物”和“固体载体”可以互换使用，其是指可以附着抗原或抗体的固体表面。对用于本发明的固相载体没有特别的限制，商品化的固相载体及任何可用于免疫分析的固相载体均可用于本发明。示例性的固相载体可以是磁珠(如羧基磁珠)、酶标板、塑料板、塑料管、乳胶珠、琼脂糖珠、玻璃、硝酸纤维素膜、尼龙膜、二氧化硅板或微芯片，但本发明不限于此。
在本发明实施方式中，可以通过以下方式产生包被在固相载体上的抗HCV抗原的抗体和HCV抗原：一方面，可以将HCV抗原的抗体和HCV抗原单独包被在不同的固相载体上，随后将经包被的固相载体混合。另一方面，也可以先将HCV抗原的抗体和HCV抗原混合，随后再与固相载体包被。
可用于本发明实施方式中的标记物是本领域技术人员所熟知的，例如可以是碱性磷酸酶(ALP)、氧化物酶、微过氧化物酶、辣根过氧化物酶、β-半乳糖苷酶、葡萄糖氧化酶以及葡萄醣6-磷酸脱氢酶等的酶；异硫氰酸荧光素、四甲基罗丹明异硫氰酸酯、荧光素、罗丹明、铕以及绿色荧光蛋白等的荧光物质；鲁米诺、异鲁米诺、菲啶鎓以及吖啶酯等的化学发光物质；NAD等辅酶；生物素； 35S、 14C、 32P、 131I以及 125I等的放射性物质，但本发明不限于此。
本领域技术人员能够依据所使用的标记物的种类选择合适的发光底物，从而产生可检测的信号。例如，当使用碱性磷酸酶作为标记物时，可采用3-(2-螺旋金刚烷)-4-甲氧基-4-(3-磷氧酰)-苯基-1,2-二氧环乙烷作为发 光底物，该底物会被碱性磷酸酶所分解，脱去一个磷酸基，生成不稳定的中间产物，该中间产物通过分子内电子转移产生间氧苯甲酸甲酯阴离子，处于激发态的间氧苯甲酸甲酯阴离子从激发态返回基态时，产生化学发光。再通过光电倍增管对反应中所产生的光子数进行测量，所产生光子的量与样本内检测物的含量成正比。
本发明实施方式适用于ELISA、化学发光、电化学发光、POCT、免疫层析法、上转换发光、下转换发光等多种方法。
在本发明的范围中，术语ROC(receiver operating characteristic)曲线是指将诊断试验结果划分为若干临界点，以每个临界点对应的灵敏度为纵坐标，特异度为横坐标，作图得到的曲线。ROC曲线是一种全面、准确评价诊断试验的有效工具。ROC曲线的另一个作用是确定检测的最佳阈值。ROC曲线法确定临界点多数情况下，选择曲线上尽量靠近左上方的点，确定临界点为最佳。在应用中，根据ROC曲线，结合各切点的灵敏度和特异度结果，选择曲线上尽量靠近左上方约登指数(Youden index)最大的切点为最佳临界点，从而使试验的灵敏度和特异度均较高，同时误诊率和漏诊率较小。
在用于制备固相试剂时，本发明实施方式的抗原例如可以多聚体、重组抗原、天然抗原、抗原片段或抗原肽的形式存在。
在用于制备固相试剂和/或标记试剂时。本发明实施方式的抗体例如可以单克隆抗体、多克隆抗体、重组抗体、嵌合抗体、人源化抗体、抗体的抗原结合片段的形式存在。
在本发明实施方式中，抗体可以源于鼠、兔、山羊、绵羊、鸡，但本发明不限于此。
在本发明实施方式中，“抗人抗体”是指可以与人抗体特异性结合的抗体。
在本发明实施方式中，标记试剂中的标记体可以经由本领域常规技术(例如，化学键结合)的方法与标记物连接。
在本发明实施方式中，通过检测值与发光阈值的比值(COI值)确定结果为阳性或阴性。例如，当比值大于等于1.1时，判断结果为阳性，表明有至少一种待测物质的检测结果为阳性。当比值介于0.9～1.1时，判断结果为灰区，既不确定为阳性或阴性；当比值小于0.9时，判断结果为阴性，表明所针对所有待测物质的检测结果均为阴性。
在本发明实施方式中，所述固相试剂和所述标记试剂中各组分的浓度被设计为使得，在同一个反应体系下采用所述固相试剂中的每一种包被在固相载体上的组分和所述标记试剂中相应带标记物的组分单独对内部参考品进行检测时，所对应的发光阈值基本上相同。例如，在确定的反应体系下，采用某一种待测物质对应的固相组分和标记组分单独对ISO 18113-1:2009中定义的内部参考品进行检测，保持标记组分的浓度为一恒定值，调节固相组分的浓度，调配发光阈值至预设值；或者，保持固相组分的浓度为一恒定值，调节标记组分的浓度，调配发光阈值至预设值。
在本发明实施方式中，“内部参考品”是指应体系组成确定的标准和依据，其具有如国际标准ISO 18113-1:2009中的定义并可依据该标准而获得。内部参考品是医疗器械生产者用于验证产品性能的样本，是产品原材料选择、制备、鉴定以及确定原材料质量标准，产品生产工艺确定，反应体系组成、反应条件等的最重要的确定标准和依据。对于定性项目，企业内部参考品的鉴别样本，是判断特定疾病、状态或被测量存在和不存在的界限的数值或量值的样本。
在本发明实施方式中，“基本上相同”是指相对偏差在±10％以内，如±5％以内、±3％以内、±2％以内或±1％以内。
如图1所示，本发明实施方式提供了可选择性检测待测样本中的HCV 抗原和/或抗HCV抗原的抗体的免疫分析仪。该免疫分析仪包括样本装置10、试剂装置20、反应装置30、光测部件40和控制装置50。该免疫分析仪还可以包括显示部件(未示出)。
样本装置10用于承载待测试的样本，吸取样本后提供给反应装置30。样本装置10包括样本存储部件11和样本分注部件12。样本存储部件11用于存储待测样本。在一些实施方式中，样本存储部件11可以包括样本分配模块(SDM，Sample Delivery Module)及前端轨道。在另一些实施方式中，样本存储部件11也可以是样本盘，样本盘包括多个可以放置诸如样本管的样本位，样本盘通过转动其盘式结构，可以将样本调度到相应位置，例如供样本分注部件12吸取样本的位置。样本分注部件12用于吸取样本并排放到待加样的反应杯中。样本分注部件12例如可以包括样本针，样本针通过二维或三维的驱动机构来在空间上进行二维或三维的运动，从而样本针可以移动去吸取样本存储部件11所承载的样本，以及移动到待加样的反应杯，并向反应杯排放样本。
试剂装置20用于承载试剂，吸取试剂后提供给反应装置30。试剂装置20包括试剂存储部件13和试剂分注部件14。试剂存储部件13用于存储试剂盒。在一些实施方式中，试剂存储部件13可以为试剂盘，试剂盘呈圆盘状结构设置，具有多个用于承载试剂容器的位置，试剂存储部件13能够转动并带动其承载的试剂容器转动，用于将试剂容器转动到特定的位置，例如被试剂分注部件14吸取试剂的位置。试剂存储部件13的数量可以为一个或多个。试剂分注部件14用于吸取试剂盒中的试剂并排放到待加试剂的反应杯中。在一些实施方式中，试剂分注部件14可以包括试剂针，试剂针通过二维或三维的驱动机构来在空间上进行二维或三维的运动，从而试剂针可以移动去吸取试剂存储部件13所承载的试剂，以及移动到待加试剂的反应杯，并向反应杯排放试剂。
其中，试剂存储部件13用于存储试剂盒。在一个实施方式中，试剂盒包括固相试剂和标记试剂。固相试剂包括第一固相组分和第二固相组分，所述第一固相组分包括包被在固相载体上的抗HCV抗原的抗体，所述第二固相组分包括包被在固相载体上的HCV抗原。标记试剂包括第一标记组分和第二标记组分，所述第一标记组分包括带标记物的抗HCV抗原的抗体，所述第二标记组分包括带标记物的抗人抗体。
反应装置30具有至少一个放置位，放置位用于放置反应杯并孵育反应杯中的反应液。例如，反应装置30可以为反应盘，其呈圆盘状结构设置，具有一个或多个用于放置反应杯的放置位，反应盘能够转动并带动其放置位中的反应杯转动，用于在反应盘内调度反应杯以及孵育反应杯中的反应液。
光测部件40用于对孵育完成的反应液进行光测定，得到样本的反应数据。例如光测部件40对待测的反应液的发光强度进行检测，通过定标曲线，计算样本中待测成分的浓度等。在一些实施方式中，光测部件40分离设置于反应装置30的外面。
免疫分析仪还包括发光底物分注装置(未示出)。该发光底物分注装置与存储发光底物的容器连接并用于将发光底物注入到待加发光底物的反应杯中。
如图2，控制装置50至少包括：处理组件51、RAM 52、ROM 53、通信接口54、存储器56和I/O接口55，其中，处理组件51、RAM 52、ROM 53、通信接口54、存储器56和I/O接口55通过总线57进行通信。
处理组件可以为CPU、GPU或其它具有运算能力的芯片。
存储器56中装有操作系统和应用程序等供处理器组件51执行的各种计算机程序及执行该计算机程序所需的数据。另外，在样本检测过程中，如有需要本地存储的数据，均可以存储到存储器56中。
I/O接口55由比如USB、IEEE1394或RS-232C等串行接口、SCSI、IDE或IEEE1284等并行接口以及由D/A转换器和A/D转换器等组成的模拟信号接口构成。I/O接口55上连接有由键盘、鼠标、触摸屏或其它控制按钮构成的输入设备，用户可以用输入设备直接向控制装置50输入数据。另外，I/O接口55上还可以连接由具有显示功能的显示器，例如：液晶屏、触摸屏、LED显示屏等，控制装置50可以将处理的数据以图像显示数据输出到显示器上进行显示，例如：分析数据、仪器运行参数等。
通信接口54是可以是目前已知的任意通信协议的接口。通信接口54通过网络与外界进行通信。控制装置50可以通过通信接口54以一定的通信协议，与通过该网连接的任意装置之间传输数据。
控制装置50配置用于接收测试指令，所述测试指令包括待测物质的类型，并且响应于所述测试指令执行下列步骤：
控制样本分注部件12将样本存储部件11中的待测样本加入到反应装置30上的反应杯中；
控制试剂分注部件14将试剂盒中与所述待测物质的类型所对应的至少一种固相组分加入到反应装置30上的反应杯中，以使所述待测样本与所加入的至少一种固相组分在所述反应杯中混合并孵育一段时间，使得固相载体上的包被体能与所述待测样本中的所述待测物质结合；
控制试剂分注部件14进一步将试剂盒中与所述待测物质的类型对应的标记组分加入所述反应杯，以使所加入的标记组分与所述反应杯中的混合物混合并孵育一段时间，使得所加入的标记组分中的标记体能与所加入的固相试剂中的包被体上结合的所述待测物质结合；
控制发光底物分注装置将发光底物加入所述反应杯中；以及
根据光测部件40中所测得发光值与发光阈值的比值获得检测结果。
通过本发明实施方式提供的免疫分析仪，能够基于一种试剂盒，根据 用户需求选择性地检测不同的待测物质，提高检测灵活性，满足用户的不同场景需求。
实验材料：
抗HCV抗原的抗体来自于Meridian Life Science；
抗人抗体来自于:Jackson ImmunoResearch；
HCV抗原来自于Meridian Life Science；
碱性磷酸酶来自于罗氏制药；
磁珠来自于Thermo Fisher。
固相包被物的制备：
首先将抗原/抗体进行预处理，通过透析去除其缓冲基质中的保护组分。按照每毫克磁珠加入1-40μg的抗原/抗体的比例(优选5-30μg)进行包被。在反应过程中磁珠表面的羧基在EDC/NHS催化下与抗原/抗体的氨基进行偶联。取20mg表面修饰有羧基的磁性微球，超声分散于10mM MES缓冲液中，加入80mg EDC和120mg NHS，超声混合均匀后，置于37℃摇床15min。之后在处理后的磁珠中，按照比例加入抗原/抗体，混匀，并置于37℃摇床反应10-18h。清洗封闭后，制备得到包被有抗原/抗体的磁性微球。选择50mM Tris pH 7.4缓冲液将包被抗原/抗体的磁性微球稀释至0.3mg/mL。
标记组分的制备：
将抗人抗体、抗HCV抗原的抗体标记信号标记物。在本发明实施例中，信号标记物为碱性磷酸酶。选择50mM MES pH 6.0缓冲液分别将抗人抗体信号标记物、抗HCV抗原的抗体信号标记物进行稀释，配制得标记组分。
检测步骤：
第一步，将样本和固相试剂添加到反应管中，在37℃孵育10分钟，使得固相表面的抗原/抗体能与样本中对应的待测物质结合。在反应管内孵育完成后，结合在固相上的物质将置于一个磁场内被吸住，结合在磁珠固相上的物质被保留，而未结合的物质被冲洗除去。
第二步，选择所需的标记组分加到反应管中孵育，进行混匀，在37℃孵育10分钟，与第一步形成的结合物结合复合物。在反应管内孵育完成后，该复合物被磁场吸住，而其他未结合的物质被清洗除去。
第三步，将AMPPD添加到反应管内，产生化学发光。再通过光电倍增管对反应所产生的光子数进行测量，以得到样本的化学发光信号值。
将发光信号值除以阈值，得到COI值。将样本的测试结果COI值与参考值(参考值为1.10)进行比较，如果大于或等于1.1，表示样本中检测物的一种或多种为阳性；如果小于0.90，表示样本中的检测物均为阴性。COI在0.90至1.10之间，结果为灰区(不确定)。
在本发明实施方式中，阴性符合率是指使用本发明实施方式的测试方法得到判断为阴性的样本个数占实际参与评估的阴性样本的比例，阳性符合率是指使用本发明实施方式的测试方法得到判断为阳性的样本个数占实际参与评估的阳性样本的比例；样本的真实阴阳性结果来自医院诊断结果。
实施例1 HCV检测的发光阈值确定
选取具有明确临床诊断结果的样本，其中HCV抗原检测样本563例(阴性样本317例、阳性样本246例)、Anti-HCV抗体检测样本600例(阴性样本320例、阳性样本280例)。
采用由“固相包被物的制备”所得到的包被有HCV抗原的磁性微球作为固相组分，由“标记组分的制备”所得到的带有碱性磷酸酶的抗人抗体作为标记组分，根据“检测步骤”测试不同发光阈值下，检测抗HCV抗原的抗体的阳性符合率和阴性符合率。类似地，采用由“固相包被物的制备”所得到的包 被有抗HCV抗原的抗体的磁性微球作为固相组分，由“标记组分的制备”所得到的带有碱性磷酸酶的抗HCV抗原的抗体作为标记组分，根据“检测步骤”测试不同发光阈值下，检测HCV抗原的阳性符合率和阴性符合率。结果汇总于表1中。
表1
Figure PCTCN2019130124-appb-000001
由表1可知，当发光阈值选择为70000左右时，总体符合率最高，将试剂盒的发光阈值选择为70000。
实施例2 HCV选择性检测的诊断效能研究
固相试剂的制备：采用由“固相包被物的制备”所得到的包被有HCV抗原的磁性微球和包被有抗HCV抗原的抗体的磁性微球作为固相试剂；
标记组分1的制备：按照“标记组分的制备”，使用制备得到带有碱性磷酸酶的抗人抗体，将其与相应的固相组分搭配测定内部参考品。选择发光阈值在70000(允许±3％偏差)对应的稀释度为标记组分1的稀释度；
标记组分2的制备：按照“标记组分的制备”，使用制备得到带有碱性磷酸酶的抗HCV抗原的抗体，将其与相应的固相组分搭配测定内部参考品。 选择发光阈值在70000(允许±3％偏差)对应的稀释度为标记组分2的稀释度。
选取具有明确临床诊断结果的样本共1000例。其中，HCV抗体和抗原阴性样本670例，HCV抗体或抗原阳性样本330例(其中HCV抗体阳性318例，其中HCV抗原阳性211例)。
当需要检测抗HCV抗原的抗体时，选择固相试剂搭配标记组分1按照“检测步骤”测得发光信号值。
当需要检测HCV抗原时，选择固相试剂搭配标记组分2按照“检测步骤”测得发光信号值。
当需要混合检测HCV抗原抗体和HCV抗原时，选择固相试剂搭配标记组分1和2搭配按照“检测步骤”测得发光信号值。
结果如下表2所示。
表2
Figure PCTCN2019130124-appb-000002
由表2可知，单独检测抗CV抗体时，阳性符合率为100％、阴性符合率为99.06％，总体符合率为99.70％；单独检测HCV抗原时，阳性符合率为100％、阴性符合率为99.53％，总体符合率为99.90％；混合检测HCV抗原和抗体，阳性符合率为100％、阴性符合率为98.48％，总体符合率为99.95％。本发明实施方式能够实现选择性地检测待测样本中的HCV抗原和/或抗HCV抗原。

Claims (14)

  1. 一种可选择性检测待测样本中的HCV抗原和/或抗HCV抗原的抗体的免疫分析仪，包括：
    样本装置，具有样本存储部件和样本分注部件，所述样本存储部件用于存储待测样本，所述样本分注部件用于吸取所述待测样本并排放到待加样的反应杯中；
    试剂装置，具有试剂存储部件和试剂分注部件，所述试剂存储部件用于存储试剂盒，所述试剂盒包括固相试剂和标记试剂，所述固相试剂包括第一固相组分和第二固相组分，所述第一固相组分包括包被在固相载体上的抗HCV抗原的抗体，所述第二固相组分包括包被在固相载体上的HCV抗原，所述标记试剂包括第一标记组分和第二标记组分，所述第一标记组分包括带标记物的抗HCV抗原的抗体，所述第二标记组分包括带标记物的抗人抗体，所述试剂分注部件用于吸取所述试剂存储部件上存储的试剂盒中的固相组分和标记组分并排放到待加试剂的反应杯中；
    发光底物分注装置，与存储发光底物的容器连接并用于将发光底物注入到待加发光底物的反应杯中；
    反应装置，具有多个用于放置所述反应杯的放置位并用于孵育所述反应杯中的反应液；
    光测部件，用于对孵育完成的反应液进行光测定，以得到待测样本的检测结果；
    控制装置，与所述样本装置、所述试剂装置、所述发光底物分注装置和所述光测部件电连接，配置用于：
    接收测试指令，所述测试指令包括待测物质的类型；
    响应于所述测试指令：
    控制所述样本分注部件将所述样本存储部件中的待测样本加入到 所述反应装置上的反应杯中；
    控制所述试剂分注部件将所述待测物质的类型所对应的至少一种固相组分加入到所述反应装置上的反应杯中，以使所述待测样本与所述至少一种固相组分在所述反应杯中混合并孵育一段时间，使得包被在固相载体上的包被体能与所述待测样本中的所述待测物质结合；
    控制所述试剂分注部件进一步将所述待测物质的类型所对应的至少一种标记组分加入所述反应杯，以使所加入的标记组分与所述反应杯中的混合物混合并孵育一段时间，使得所加入的标记组分中的标记体能与包被在固相载体上的包被体上结合的所述待测物质结合，形成包被体-待测物质-标记体的复合物；
    控制所述发光底物分注装置将发光底物加入所述反应杯中；以及
    根据所述光测部件中所测得发光值与发光阈值的比值获得检测结果。
  2. 权利要求1所述的免疫分析仪，其中，在所述固相试剂中，包被在固相载体上的抗HCV抗原的抗体和HCV抗原以单独分装的形式存在；
    在所述标记试剂中，带标记物的抗HCV抗原的抗体和抗人抗体以单独分装或预先混合的形式存在。
  3. 权利要求1所述的免疫分析仪，其中，在所述固相试剂中，包被在固相载体上的抗HCV抗原的抗体和HCV抗原以预先混合的形式存在；
    在所述标记试剂中，带标记物的抗HCV抗原的抗体和抗人抗体以单独分装的形式存在。
  4. 权利要求1至3中任一项所述的免疫分析仪，其中，所述固相试剂和所述标记试剂中各物质的浓度被设计为使得，在同一个反应体系下采用所述固相试剂中的每一种包被在固相载体上的包被体和所述标记试剂中对应带标记物的标记体单独对内部参考品进行检测时，所对应的发光阈值基 本上相同。
  5. 权利要求1至4中任一项所述的免疫分析仪，其中，所述抗人抗体选自抗人IgG抗体和抗人IgM抗体中的至少一种。
  6. 一种检测HCV的免疫分析方法，包括以下步骤：
    将待测样本与包被在固相载体上的抗HCV抗原的抗体和HCV抗原混合并孵育一段时间，使得包被在固相载体上的抗HCV抗原的抗体和HCV抗原能与所述待测样本中的待测物质结合；
    对所述待测样本和所述固相试剂的混合物进行清洗，除去未结合的物质；
    向经清洗的混合物中加入带标记物的抗HCV抗原的抗体和带标记物的抗人抗体并孵育，使得带标记物的抗HCV抗原的抗体和带标记物的抗人抗体能分别与包被在固相载体上的抗HCV抗原的抗体和HCV抗原上结合的所述待测物质结合，形成复合物；
    对所述复合物进行清洗，除去未结合的物质；
    在经清洗的复合物中加入发光底物，以检测所述待测样本中抗HCV抗原和HCV抗原的混合检测值。
  7. 权利要求6所述的方法，其中，包被在固相载体上的抗HCV抗原的抗体和HCV抗原被分别加入到所述待测样本中进行混合并孵育，或者以预先混合的形式被加入到所述待测样本中进行混合并孵育。
  8. 权利要求6或7所述的方法，其中，带有标记物的抗HCV抗原的抗体和抗人抗体被分别加入到所述经清洗的混合物中进行混合并孵育，或者以预先混合的形式被加入到所述经清洗的混合物中进行混合并孵育。
  9. 权利要求6至8中任一项所述的方法，其中，所述固相试剂和所述标记试剂中各物质的浓度被设计为使得，在同一个反应体系下采用所述固相试剂中的每一种包被在固相载体上的包被体和所述标记试剂中对应带标 记物的标记体单独对内部参考品进行检测时，所对应的发光阈值基本上相同。
  10. 一种检测HCV的试剂盒，包括：
    固相试剂，包含包被在固相载体上的抗HCV抗原的抗体和HCV抗原；
    标记试剂，包含带标记物的抗HCV抗原的抗体和抗人抗体。
  11. 权利要求10所述的试剂盒，其中，所述抗人抗体选自抗人IgG抗体和抗人IgM抗体中的至少一种。
  12. 权利要求10或11中所述的试剂盒，其中，所述固相试剂和所述标记试剂中各物质的浓度被设计为使得，在同一个反应体系下采用所述固相试剂中的每一种包被在固相载体上的包被体和所述标记试剂中对应带标记物的标记体单独对内部参考品进行检测时，所对应的发光阈值基本上相同。
  13. 权利要求10至12中任一项所述的试剂盒，其中，在所述固相试剂中，包被在固相载体上的抗HCV抗原的抗体和HCV抗原以单独分装或预先混合的形式存在。
  14. 权利要求10至13中任一项所述的试剂盒，其中，在所述标记试剂中，带有标记物的抗HCV抗原的抗体和抗人抗体以单独分装或预先混合的形式存在。
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Citations (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH02236455A (ja) * 1989-03-10 1990-09-19 Jeol Ltd 自動免疫測定装置の発光試薬注入装置
CN1378085A (zh) * 2001-03-28 2002-11-06 奥索临床诊断有限公司 用于感染早期检测的丙型肝炎抗原－抗体联合测定
JP2003329696A (ja) * 2002-05-15 2003-11-19 Horiba Ltd 化学発光酵素免疫測定装置
WO2004070387A1 (en) * 2003-02-04 2004-08-19 Abbott Laboratories Improved method of detection of hcv antibodies in combination assay or sole antibody assay
CN1869702A (zh) * 2005-05-24 2006-11-29 穆海东 肝炎与肝硬化的集成检测反应板和蛋白芯片试剂盒
CN105004862A (zh) * 2015-07-06 2015-10-28 山东博科生物产业有限公司 一种高通量联合检测丙肝病毒抗原抗体的试剂
CN206540925U (zh) * 2017-03-09 2017-10-03 厦门先明生物技术有限公司 一种集成试剂组线形推进并行处理反应装置
CN108226482A (zh) * 2012-03-26 2018-06-29 埃克塞尔柯尔有限责任公司 用于检测分析物的装置和方法
CN208721691U (zh) * 2018-09-07 2019-04-09 四川沃文特生物技术有限公司 一种发光化学免疫分析仪

Patent Citations (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH02236455A (ja) * 1989-03-10 1990-09-19 Jeol Ltd 自動免疫測定装置の発光試薬注入装置
CN1378085A (zh) * 2001-03-28 2002-11-06 奥索临床诊断有限公司 用于感染早期检测的丙型肝炎抗原－抗体联合测定
JP2003329696A (ja) * 2002-05-15 2003-11-19 Horiba Ltd 化学発光酵素免疫測定装置
WO2004070387A1 (en) * 2003-02-04 2004-08-19 Abbott Laboratories Improved method of detection of hcv antibodies in combination assay or sole antibody assay
CN1869702A (zh) * 2005-05-24 2006-11-29 穆海东 肝炎与肝硬化的集成检测反应板和蛋白芯片试剂盒
CN108226482A (zh) * 2012-03-26 2018-06-29 埃克塞尔柯尔有限责任公司 用于检测分析物的装置和方法
CN105004862A (zh) * 2015-07-06 2015-10-28 山东博科生物产业有限公司 一种高通量联合检测丙肝病毒抗原抗体的试剂
CN206540925U (zh) * 2017-03-09 2017-10-03 厦门先明生物技术有限公司 一种集成试剂组线形推进并行处理反应装置
CN208721691U (zh) * 2018-09-07 2019-04-09 四川沃文特生物技术有限公司 一种发光化学免疫分析仪

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