CN109001468A - A kind of detection kit of CMV antibody IgM - Google Patents

A kind of detection kit of CMV antibody IgM Download PDF

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Publication number
CN109001468A
CN109001468A CN201810883445.7A CN201810883445A CN109001468A CN 109001468 A CN109001468 A CN 109001468A CN 201810883445 A CN201810883445 A CN 201810883445A CN 109001468 A CN109001468 A CN 109001468A
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added
calibration object
microballon
reagent
herpes simplex
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Pending
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CN201810883445.7A
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Chinese (zh)
Inventor
宣生良
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One Drop Biotechnology (changzhou) Co Ltd
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One Drop Biotechnology (changzhou) Co Ltd
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Priority to CN201810883445.7A priority Critical patent/CN109001468A/en
Priority to CN202310698886.0A priority patent/CN116500279A/en
Publication of CN109001468A publication Critical patent/CN109001468A/en
Pending legal-status Critical Current

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    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/569Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
    • G01N33/56983Viruses
    • G01N33/56994Herpetoviridae, e.g. cytomegalovirus, Epstein-Barr virus
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/54313Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being characterised by its particulate form
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/569Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
    • G01N33/56983Viruses
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/58Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances
    • G01N33/582Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances with fluorescent label
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/6854Immunoglobulins
    • YGENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02ATECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Abstract

The invention discloses a kind of detection kits of the antibody IgMs such as cytomegalovirus, belong to detection reagent technical field.It includes compounded microbeads suspension, fluorescence display agent, Sample dilution and cleaning solution, production technology saves liquid according to microballon is prepared, the activation and coating of microballoon, it prepares fluorescent marker and prepares concentrated cleaning solution and produced, and combine calibration object 1, calibration object 2, calibration object 3, the specific antigen of reference microspheres and NSC microballoon reference substance, it solves when the toxoplasma in one sample of detection, rubeola, when cytomegalovirus and herpes simplex virus type 1 and herpes simplex virus type 2, it needs individually with four kinds of detection reagents respectively to toxoplasma, rubeola, cytomegalovirus, the problem of herpes simplex virus type 1 and herpes simplex virus type 2 are detected, shorten detection time, to improve detection efficiency.

Description

A kind of detection kit of CMV antibody IgM
Technical field
The present invention relates to a kind of detection kits of the antibody IgMs such as cytomegalovirus, belong to detection reagent technical field, this Invention is that one kind can be in a micropore, while measuring the toxoplasma in a sample, rubella virus, and cytomegalovirus is single Pure 1 type of herpesviral, the detection kit of herpes simplex virus type 2 IgM antibody.
Background technique
Toxoplasma, rubeola, cytomegalovirus, herpes simplex virus type 1, herpes simplex virus type 2 are that today's society is common Virus, however as the progress of medical level, toxoplasma, rubeola, cytomegalovirus and herpes simplex virus type 1 and simple blister 2 type of exanthema virus can be transferred through existing detection reagent and be detected, and then do further treatment again, however when one sample of detection When toxoplasma, rubeola, cytomegalovirus and herpes simplex virus type 1 and herpes simplex virus type 2 in this, need individually to use Four kinds of detection reagents respectively to toxoplasma, rubeola, cytomegalovirus and herpes simplex virus type 1 and herpes simplex virus type 2 into Row detection, increases detection time, reduces detection efficiency.
Summary of the invention
Technical problem to be solved by the present invention lies in: a kind of detection kit of the antibody IgMs such as cytomegalovirus is provided, It is solved when toxoplasma, rubeola, cytomegalovirus, herpes simplex virus type 1 and the herpe simplex disease in one sample of detection When malicious 2 type, need individually with four kinds of detection reagents respectively to toxoplasma, rubeola, cytomegalovirus, herpes simplex virus type 1 The problem of being detected with herpes simplex virus type 2.
The technical problems to be solved by the invention take following technical scheme to realize:
A kind of detection kit of the antibody IgMs such as cytomegalovirus, detection kit includes compounded microbeads suspension, fluorescence display Agent, Sample dilution and cleaning solution,
(1) compounded microbeads suspension: including cognizable polystyrene microbeads in suspension, and polystyrene microbeads are coated with respectively There is following antigen: toxoplasma antigen, rubella antigens, big and small extracellular antigen, -1 antigen of herpes simplex virus and herpes simplex virus-2 Antigen;
(2) fluorescence display agent: the sheep Anti-Human's IgM(specificity γ marked using the phycoerythrin that initial concentration is 1.0 ug/mL Chain), dilution calibration is working solution concentration 0.6ug/mL, 2-8 DEG C of storage;
(3) Sample dilution: phosphate-containing buffer is used for diluted sample;
(4) cleaning solution: phosphate-containing buffer need to be used with 10 times of purified water dilutions, board-washing.
As preferred embodiment, the production technology of kit mainly has the following steps:
1. preparing microballon saves liquid, microballon saves the NaCl reagent that liquid main component includes: i.e. 8.0g/L, 2.9g/L's Na2HPO412H2O reagent, the KCl reagent of 2.4g/L, the KH2PO4 reagent of 2.4g/L, the Tris reagent of 0.5g/L, 0.5mL/ The Tween20 reagent of L, the Sodium Azide reagent of 0.5g/L.
2. the activation and coating of microballoon take 1mL particular number microballon that 1.5mL centrifuge tube is added, 5-50uL EDC(1- second is added Base-(3- dimethylaminopropyl) phosphinylidyne diimmonium salt hydrochlorate) and 5-50uL SuLfo-NHS(N- hydroxy succinyl Asia Amine), 10-100ug specific antigen is added, is vortexed and mixes 30 seconds, ultrasound mixes 30 seconds, and 2-8 DEG C shakes up overnight.
3. fluorescent marker: the sheep Anti-Human IgM(specificity γ chain of phycoerythrin label), 1 μ g/mL of initial concentration, use is micro- Pearl saves liquid and is diluted to working concentration, and working concentration is 0.6 μ g/L.
4. concentrated cleaning solution: it is consistent to save the ingredient of liquid for ingredient and microballon in concentrated cleaning solution, and concentrated cleaning solution respectively at Dividing concentration is 10 times that microballon saves each constituent concentration of liquid.
As preferred embodiment, specific antigen includes calibration object 1, calibration object 2, calibration object 3, reference microspheres and NSC microballoon reference substance, The people IgM of 2-5ug/mL is added in calibration object 1, and the people IgM of 6-10ug/mL is added in calibration object 2, and 12-18ug/mL is added in calibration object 3 People IgM, reference microspheres be added 8-15ug/mL anti-human IgM, NSC microballoon as control, be added without any antigen.
The beneficial effects of the present invention are: the present invention includes compounded microbeads suspension, fluorescence display agent, Sample dilution and washes Liquid is washed, production technology saves liquid, the activation of microballoon and coating, prepares fluorescent marker and prepare thickening and washing according to microballon is prepared Liquid is produced, and combine calibration object 1, calibration object 2, calibration object 3, reference microspheres and NSC microballoon reference substance specific antigen, solution It has determined when detecting the toxoplasma in a sample, rubeola, cytomegalovirus and herpes simplex virus, has needed individually with four kinds Detection reagent respectively examines toxoplasma, rubeola, cytomegalovirus and herpes simplex virus type 1 and herpes simplex virus type 2 The problem of survey, shortens detection time, to improve detection efficiency.
Specific embodiment
In order to be easy to understand to technical means, creative features, achievable purpose and effectiveness of the invention, below with reference to tool Body embodiment, the present invention is further explained.
A kind of detection kit of the antibody IgMs such as cytomegalovirus, detection kit includes compounded microbeads suspension, fluorescence Developer, Sample dilution and cleaning solution,
(1) compounded microbeads suspension: including cognizable polystyrene microbeads in suspension, and polystyrene microbeads are coated with respectively There is following antigen: toxoplasma antigen, rubella antigens, big and small extracellular antigen, -1 antigen of herpes simplex virus and herpes simplex virus-2 Antigen;
(2) fluorescence display agent: the sheep Anti-Human's IgM(specificity γ marked using the phycoerythrin that initial concentration is 1.0 ug/mL Chain), dilution calibration is working solution concentration 0.6ug/mL, 2-8 DEG C of storage;
(3) Sample dilution: phosphate-containing buffer is used for diluted sample;
(4) cleaning solution: phosphate-containing buffer need to be used with 10 times of purified water dilutions, board-washing.
The production technology of kit mainly has the following steps:
1. preparing microballon saves liquid, microballon saves the NaCl reagent that liquid main component includes: i.e. 8.0g/L, 2.9g/L's Na2HPO412H2O reagent, the KCl reagent of 2.4g/L, the KH2PO4 reagent of 2.4g/L, the Tris reagent of 0.5g/L, 0.5mL/ The Tween20 reagent of L, the Sodium Azide reagent of 0.5g/L.
2. the activation and coating of microballoon take 1mL particular number microballon that 1.5mL centrifuge tube is added, 5-50uL EDC(1- second is added Base-(3- dimethylaminopropyl) phosphinylidyne diimmonium salt hydrochlorate) and 5-50uL SuLfo-NHS(N- hydroxy succinyl Asia Amine), 10-100ug specific antigen is added, is vortexed and mixes 30 seconds, ultrasound mixes 30 seconds, and 2-8 DEG C shakes up overnight.
When producing detection kit, needs to place the shaking table of 4 kinds of antigen-antibody bottles simultaneously, be coated with different numbers respectively Microballoon, steps are as follows:
1) wash: use filtering and washing, it is washed after microballon be transferred to microballon save liquid;
2) verification quality Quality Control: is come by using Quality Control disk;
3) a variety of microballons that have been coated with are mixed, saves liquid with a small amount of microballon and wash lower bottle wall residual microballon, and mix, finally with micro- Pearl saves liquid constant volume, dispenses after the assay was approved.
3. fluorescent marker: the sheep Anti-Human IgM(specificity γ chain of phycoerythrin label), 1 μ g/mL of initial concentration, use is micro- Pearl saves liquid and is diluted to working concentration, and working concentration is 0.6 μ g/L.
4. concentrated cleaning solution: microballon saves 10 times of concentration versions of formula of liquid, and ingredient and microballon save liquid in concentrated cleaning solution Ingredient it is consistent, and each constituent concentration of concentrated cleaning solution is that microballon saves 10 times of each constituent concentration of liquid.
Specific antigen includes calibration object 1, calibration object 2, calibration object 3, reference microspheres and NSC microballoon reference substance, and 2- is added in calibration object 1 The people IgM of 6-10ug/mL is added in the people IgM of 5ug/mL, calibration object 2, and the people IgM of 12-18ug/mL is added in calibration object 3, and reference is micro- The anti-human IgM of 8-15ug/mL is added in ball, and NSC microballoon is added without any antigen as control.
The above shows and describes the basic principles and main features of the present invention and the advantages of the present invention.The technology of the industry Personnel are it should be appreciated that the present invention is not limited to the above embodiments, without departing from the spirit and scope of the present invention, this hair Bright to will also have various changes and improvements, these changes and improvements all fall within the protetion scope of the claimed invention.The present invention claims Protection scope is defined by the appending claims and its equivalent thereof.

Claims (3)

1. a kind of detection kit of the antibody IgMs such as cytomegalovirus, it is characterised in that: detection kit includes that compounded microbeads are outstanding Supernatant liquid, fluorescence display agent, Sample dilution and cleaning solution,
Compounded microbeads suspension: including cognizable polystyrene microbeads in suspension, and polystyrene microbeads are coated with respectively Following antigen: toxoplasma antigen, rubella antigens, big and small extracellular antigen, -1 antigen of herpes simplex virus and herpes simplex virus-2 are anti- It is former;
Fluorescence display agent: the sheep Anti-Human IgM(specificity γ chain marked using the phycoerythrin that initial concentration is 1.0 ug/mL), Dilution calibration is working solution concentration 0.6ug/mL, 2-8 DEG C of storage;
Sample dilution: phosphate-containing buffer is used for diluted sample;
Cleaning solution: phosphate-containing buffer need to be used with 10 times of purified water dilutions, board-washing.
2. the detection kit of the antibody IgMs such as a kind of cytomegalovirus according to claim 1, it is characterised in that: kit Production technology mainly have the following steps:
It prepares microballon and saves liquid, microballon saves the NaCl reagent that liquid main component includes: i.e. 8.0g/L, 2.9g/L's Na2HPO412H2O reagent, the KCl reagent of 2.4g/L, the KH2PO4 reagent of 2.4g/L, the Tris reagent of 0.5g/L, 0.5mL/ The Tween20 reagent of L, the Sodium Azide reagent of 0.5g/L;
The activation and coating of microballoon take 1mL particular number microballon that 1.5mL centrifuge tube is added, and 5-50uL EDC(1- ethyl-is added (3- dimethylaminopropyl) phosphinylidyne diimmonium salt hydrochlorate) and 5-50uL SuLfo-NHS(N- hydroxy thiosuccinimide), 10-100ug specific antigen is added, is vortexed and mixes 30 seconds, ultrasound mixes 30 seconds, and 2-8 DEG C shakes up overnight;
Fluorescent marker: the sheep Anti-Human IgM(specificity γ chain of phycoerythrin label), 1 μ g/mL of initial concentration is saved with microballon Liquid is diluted to working concentration, and working concentration is 0.6 μ g/L;
Concentrated cleaning solution: ingredient is consistent with the microballon preservation ingredient of liquid in concentrated cleaning solution, and each constituent concentration of concentrated cleaning solution 10 times for saving each constituent concentration of liquid for microballon.
3. the detection kit of the antibody IgMs such as a kind of cytomegalovirus according to claim 2, it is characterised in that: specific anti- Original includes calibration object 1, calibration object 2, calibration object 3, reference microspheres and NSC microballoon reference substance, and the people of 2-5ug/mL is added in calibration object 1 The people IgM of 6-10ug/mL is added in IgM, calibration object 2, and the people IgM of 12-18ug/mL is added in calibration object 3, and 8- is added in reference microspheres The anti-human IgM of 15ug/mL, NSC microballoon are added without any antigen as control.
CN201810883445.7A 2018-08-06 2018-08-06 A kind of detection kit of CMV antibody IgM Pending CN109001468A (en)

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CN201810883445.7A CN109001468A (en) 2018-08-06 2018-08-06 A kind of detection kit of CMV antibody IgM
CN202310698886.0A CN116500279A (en) 2018-08-06 2018-08-06 Detection kit for cytomegalovirus antibody IgM and application

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Cited By (3)

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CN114113626A (en) * 2021-10-25 2022-03-01 江苏纳迪芯生命科技研究院有限公司 Cytokine detection kit based on magnetic particle luminescence method
CN114113627A (en) * 2021-10-25 2022-03-01 江苏纳迪芯生命科技研究院有限公司 Alzheimer's disease detection kit based on magnetic particle luminescence method
CN114113590A (en) * 2021-10-25 2022-03-01 江苏纳迪芯生命科技研究院有限公司 Mycoplasma pneumoniae and chlamydia antibody detection kit based on magnetic particle luminescence method

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CN114113626A (en) * 2021-10-25 2022-03-01 江苏纳迪芯生命科技研究院有限公司 Cytokine detection kit based on magnetic particle luminescence method
CN114113627A (en) * 2021-10-25 2022-03-01 江苏纳迪芯生命科技研究院有限公司 Alzheimer's disease detection kit based on magnetic particle luminescence method
CN114113590A (en) * 2021-10-25 2022-03-01 江苏纳迪芯生命科技研究院有限公司 Mycoplasma pneumoniae and chlamydia antibody detection kit based on magnetic particle luminescence method

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RJ01 Rejection of invention patent application after publication
RJ01 Rejection of invention patent application after publication

Application publication date: 20181214