CN117347616A - 稀释剂、捕获试剂、检测试剂盒和测定方法 - Google Patents
稀释剂、捕获试剂、检测试剂盒和测定方法 Download PDFInfo
- Publication number
- CN117347616A CN117347616A CN202210734085.0A CN202210734085A CN117347616A CN 117347616 A CN117347616 A CN 117347616A CN 202210734085 A CN202210734085 A CN 202210734085A CN 117347616 A CN117347616 A CN 117347616A
- Authority
- CN
- China
- Prior art keywords
- diluent
- capture reagent
- analyte
- conductivity
- salt
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 239000003085 diluting agent Substances 0.000 title claims abstract description 107
- 239000003153 chemical reaction reagent Substances 0.000 title claims abstract description 61
- 238000001514 detection method Methods 0.000 title claims abstract description 36
- 238000003556 assay Methods 0.000 title abstract description 9
- 239000011324 bead Substances 0.000 claims description 68
- 239000012491 analyte Substances 0.000 claims description 45
- 239000003795 chemical substances by application Substances 0.000 claims description 26
- 238000002372 labelling Methods 0.000 claims description 24
- 150000003839 salts Chemical class 0.000 claims description 21
- -1 urinary microalbumin Proteins 0.000 claims description 17
- 239000000126 substance Substances 0.000 claims description 15
- 239000000872 buffer Substances 0.000 claims description 12
- 238000000034 method Methods 0.000 claims description 12
- 239000007987 MES buffer Substances 0.000 claims description 10
- 108010063628 acarboxyprothrombin Proteins 0.000 claims description 10
- 101800000407 Brain natriuretic peptide 32 Proteins 0.000 claims description 9
- 102400000667 Brain natriuretic peptide 32 Human genes 0.000 claims description 9
- 101800002247 Brain natriuretic peptide 45 Proteins 0.000 claims description 9
- 238000003018 immunoassay Methods 0.000 claims description 9
- HPNRHPKXQZSDFX-OAQDCNSJSA-N nesiritide Chemical group C([C@H]1C(=O)NCC(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@H](C(N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)NCC(=O)N[C@@H](CC(C)C)C(=O)NCC(=O)N[C@@H](CSSC[C@@H](C(=O)N1)NC(=O)CNC(=O)[C@H](CO)NC(=O)CNC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H](NC(=O)[C@H](CCSC)NC(=O)[C@H](CCCCN)NC(=O)[C@H]1N(CCC1)C(=O)[C@@H](N)CO)C(C)C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=1N=CNC=1)C(O)=O)=O)[C@@H](C)CC)C1=CC=CC=C1 HPNRHPKXQZSDFX-OAQDCNSJSA-N 0.000 claims description 9
- 238000002360 preparation method Methods 0.000 claims description 8
- 238000012360 testing method Methods 0.000 claims description 8
- 241001678559 COVID-19 virus Species 0.000 claims description 7
- 239000000427 antigen Substances 0.000 claims description 5
- 108091007433 antigens Proteins 0.000 claims description 5
- 102000036639 antigens Human genes 0.000 claims description 5
- 239000003607 modifier Substances 0.000 claims description 5
- 102000036675 Myoglobin Human genes 0.000 claims description 4
- 108010062374 Myoglobin Proteins 0.000 claims description 4
- JKMHFZQWWAIEOD-UHFFFAOYSA-N 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid Chemical compound OCC[NH+]1CCN(CCS([O-])(=O)=O)CC1 JKMHFZQWWAIEOD-UHFFFAOYSA-N 0.000 claims description 3
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 claims description 3
- 108010074051 C-Reactive Protein Proteins 0.000 claims description 3
- 102100032752 C-reactive protein Human genes 0.000 claims description 3
- 102000004420 Creatine Kinase Human genes 0.000 claims description 3
- 108010042126 Creatine kinase Proteins 0.000 claims description 3
- 102000012192 Cystatin C Human genes 0.000 claims description 3
- 108010061642 Cystatin C Proteins 0.000 claims description 3
- 239000003154 D dimer Substances 0.000 claims description 3
- 239000007995 HEPES buffer Substances 0.000 claims description 3
- 101001023833 Homo sapiens Neutrophil gelatinase-associated lipocalin Proteins 0.000 claims description 3
- 102000004889 Interleukin-6 Human genes 0.000 claims description 3
- 108090001005 Interleukin-6 Proteins 0.000 claims description 3
- 108010044467 Isoenzymes Proteins 0.000 claims description 3
- 239000007993 MOPS buffer Substances 0.000 claims description 3
- 108010048233 Procalcitonin Proteins 0.000 claims description 3
- 108700028909 Serum Amyloid A Proteins 0.000 claims description 3
- 102000054727 Serum Amyloid A Human genes 0.000 claims description 3
- 102000013394 Troponin I Human genes 0.000 claims description 3
- 108010065729 Troponin I Proteins 0.000 claims description 3
- 102100036859 Troponin I, cardiac muscle Human genes 0.000 claims description 3
- 101710128251 Troponin I, cardiac muscle Proteins 0.000 claims description 3
- 102000004987 Troponin T Human genes 0.000 claims description 3
- 108090001108 Troponin T Proteins 0.000 claims description 3
- 102000015736 beta 2-Microglobulin Human genes 0.000 claims description 3
- 108010081355 beta 2-Microglobulin Proteins 0.000 claims description 3
- 108010052295 fibrin fragment D Proteins 0.000 claims description 3
- 102000047202 human LCN2 Human genes 0.000 claims description 3
- 229940100601 interleukin-6 Drugs 0.000 claims description 3
- 239000008363 phosphate buffer Substances 0.000 claims description 3
- 239000002243 precursor Substances 0.000 claims description 3
- CWCXERYKLSEGEZ-KDKHKZEGSA-N procalcitonin Chemical compound C([C@@H](C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@H](C(=O)N[C@@H](C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)NCC(=O)N[C@@H](C(C)C)C(=O)NCC(=O)N[C@@H](C)C(=O)N1[C@@H](CCC1)C(=O)NCC(O)=O)[C@@H](C)O)NC(=O)[C@@H](NC(=O)[C@H](CC=1NC=NC=1)NC(=O)[C@H](CC=1C=CC=CC=1)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC=1C=CC=CC=1)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H](NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@@H](NC(=O)CNC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCSC)NC(=O)[C@H]1NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](CO)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC(N)=O)NC(=O)CNC(=O)[C@@H](N)CSSC1)[C@@H](C)O)[C@@H](C)O)[C@@H](C)O)C1=CC=CC=C1 CWCXERYKLSEGEZ-KDKHKZEGSA-N 0.000 claims description 3
- 102000029752 retinol binding Human genes 0.000 claims description 3
- 108091000053 retinol binding Proteins 0.000 claims description 3
- 230000002485 urinary effect Effects 0.000 claims description 3
- 101710141454 Nucleoprotein Proteins 0.000 claims description 2
- 150000003863 ammonium salts Chemical class 0.000 claims description 2
- 159000000007 calcium salts Chemical class 0.000 claims description 2
- 159000000003 magnesium salts Chemical class 0.000 claims description 2
- XAEFZNCEHLXOMS-UHFFFAOYSA-M potassium benzoate Chemical compound [K+].[O-]C(=O)C1=CC=CC=C1 XAEFZNCEHLXOMS-UHFFFAOYSA-M 0.000 claims description 2
- 159000000000 sodium salts Chemical class 0.000 claims description 2
- 230000035945 sensitivity Effects 0.000 abstract description 10
- 239000006172 buffering agent Substances 0.000 abstract description 2
- 239000000243 solution Substances 0.000 description 46
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical group [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 43
- 239000011248 coating agent Substances 0.000 description 40
- 238000000576 coating method Methods 0.000 description 40
- 238000002156 mixing Methods 0.000 description 31
- 239000000523 sample Substances 0.000 description 23
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 description 22
- 239000011780 sodium chloride Substances 0.000 description 21
- 238000007885 magnetic separation Methods 0.000 description 17
- RXNXLAHQOVLMIE-UHFFFAOYSA-N phenyl 10-methylacridin-10-ium-9-carboxylate Chemical class C12=CC=CC=C2[N+](C)=C2C=CC=CC2=C1C(=O)OC1=CC=CC=C1 RXNXLAHQOVLMIE-UHFFFAOYSA-N 0.000 description 17
- 239000012224 working solution Substances 0.000 description 17
- 238000010790 dilution Methods 0.000 description 16
- 239000012895 dilution Substances 0.000 description 16
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 12
- DZBUGLKDJFMEHC-UHFFFAOYSA-N acridine Chemical class C1=CC=CC2=CC3=CC=CC=C3N=C21 DZBUGLKDJFMEHC-UHFFFAOYSA-N 0.000 description 12
- 229960002685 biotin Drugs 0.000 description 11
- 235000020958 biotin Nutrition 0.000 description 11
- 239000011616 biotin Substances 0.000 description 11
- 239000003550 marker Substances 0.000 description 11
- 239000007853 buffer solution Substances 0.000 description 10
- 238000006243 chemical reaction Methods 0.000 description 10
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 9
- 102000012288 Phosphopyruvate Hydratase Human genes 0.000 description 9
- 108010022181 Phosphopyruvate Hydratase Proteins 0.000 description 9
- 239000000203 mixture Substances 0.000 description 9
- 239000012141 concentrate Substances 0.000 description 8
- 229920001213 Polysorbate 20 Polymers 0.000 description 7
- 229910052751 metal Inorganic materials 0.000 description 7
- 239000002184 metal Substances 0.000 description 7
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 description 7
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 description 7
- 239000004471 Glycine Substances 0.000 description 6
- OAKJQQAXSVQMHS-UHFFFAOYSA-N Hydrazine Chemical compound NN OAKJQQAXSVQMHS-UHFFFAOYSA-N 0.000 description 6
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 6
- 201000010099 disease Diseases 0.000 description 5
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 5
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 4
- 239000007983 Tris buffer Substances 0.000 description 4
- 238000004132 cross linking Methods 0.000 description 4
- 150000002739 metals Chemical class 0.000 description 4
- 239000003761 preservation solution Substances 0.000 description 4
- 239000003755 preservative agent Substances 0.000 description 4
- 230000002335 preservative effect Effects 0.000 description 4
- 235000018102 proteins Nutrition 0.000 description 4
- 102000004169 proteins and genes Human genes 0.000 description 4
- 108090000623 proteins and genes Proteins 0.000 description 4
- 238000007789 sealing Methods 0.000 description 4
- 239000004094 surface-active agent Substances 0.000 description 4
- 239000000725 suspension Substances 0.000 description 4
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 4
- 238000005406 washing Methods 0.000 description 4
- 108010022366 Carcinoembryonic Antigen Proteins 0.000 description 3
- 102100025475 Carcinoembryonic antigen-related cell adhesion molecule 5 Human genes 0.000 description 3
- 108010090804 Streptavidin Proteins 0.000 description 3
- DTQVDTLACAAQTR-UHFFFAOYSA-M Trifluoroacetate Chemical compound [O-]C(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-M 0.000 description 3
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical class OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 description 3
- 150000001875 compounds Chemical class 0.000 description 3
- 238000007865 diluting Methods 0.000 description 3
- 239000003446 ligand Substances 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 229910052755 nonmetal Inorganic materials 0.000 description 3
- 108091023037 Aptamer Proteins 0.000 description 2
- 108090001008 Avidin Proteins 0.000 description 2
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 108060003951 Immunoglobulin Proteins 0.000 description 2
- 108010058846 Ovalbumin Proteins 0.000 description 2
- 108010039918 Polylysine Proteins 0.000 description 2
- 239000006180 TBST buffer Substances 0.000 description 2
- 108010034949 Thyroglobulin Proteins 0.000 description 2
- 102000009843 Thyroglobulin Human genes 0.000 description 2
- 239000003929 acidic solution Substances 0.000 description 2
- ORILYTVJVMAKLC-UHFFFAOYSA-N adamantane Chemical compound C1C(C2)CC3CC1CC2C3 ORILYTVJVMAKLC-UHFFFAOYSA-N 0.000 description 2
- 230000001588 bifunctional effect Effects 0.000 description 2
- 230000000903 blocking effect Effects 0.000 description 2
- 229940098773 bovine serum albumin Drugs 0.000 description 2
- 239000005018 casein Substances 0.000 description 2
- 235000021240 caseins Nutrition 0.000 description 2
- 239000002131 composite material Substances 0.000 description 2
- 238000005859 coupling reaction Methods 0.000 description 2
- 239000003431 cross linking reagent Substances 0.000 description 2
- 238000003745 diagnosis Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 230000005284 excitation Effects 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 239000000706 filtrate Substances 0.000 description 2
- 125000000524 functional group Chemical group 0.000 description 2
- 102000018358 immunoglobulin Human genes 0.000 description 2
- 108010045069 keyhole-limpet hemocyanin Proteins 0.000 description 2
- 239000002923 metal particle Substances 0.000 description 2
- 229940092253 ovalbumin Drugs 0.000 description 2
- 229920000656 polylysine Polymers 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 108020003175 receptors Proteins 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 229960002175 thyroglobulin Drugs 0.000 description 2
- ZMRMMAOBSFSXLN-UHFFFAOYSA-N 4-[4-(2,5-dioxopyrrol-1-yl)phenyl]butanehydrazide Chemical compound C1=CC(CCCC(=O)NN)=CC=C1N1C(=O)C=CC1=O ZMRMMAOBSFSXLN-UHFFFAOYSA-N 0.000 description 1
- HUDPLKWXRLNSPC-UHFFFAOYSA-N 4-aminophthalhydrazide Chemical compound O=C1NNC(=O)C=2C1=CC(N)=CC=2 HUDPLKWXRLNSPC-UHFFFAOYSA-N 0.000 description 1
- 102100031126 6-phosphogluconolactonase Human genes 0.000 description 1
- 108010029731 6-phosphogluconolactonase Proteins 0.000 description 1
- 102000002260 Alkaline Phosphatase Human genes 0.000 description 1
- 108020004774 Alkaline Phosphatase Proteins 0.000 description 1
- 108010031480 Artificial Receptors Proteins 0.000 description 1
- 102100026189 Beta-galactosidase Human genes 0.000 description 1
- LTMHDMANZUZIPE-AMTYYWEZSA-N Digoxin Natural products O([C@H]1[C@H](C)O[C@H](O[C@@H]2C[C@@H]3[C@@](C)([C@@H]4[C@H]([C@]5(O)[C@](C)([C@H](O)C4)[C@H](C4=CC(=O)OC4)CC5)CC3)CC2)C[C@@H]1O)[C@H]1O[C@H](C)[C@@H](O[C@H]2O[C@@H](C)[C@H](O)[C@@H](O)C2)[C@@H](O)C1 LTMHDMANZUZIPE-AMTYYWEZSA-N 0.000 description 1
- 108010018962 Glucosephosphate Dehydrogenase Proteins 0.000 description 1
- 108010001336 Horseradish Peroxidase Proteins 0.000 description 1
- 108091034117 Oligonucleotide Proteins 0.000 description 1
- 108010094028 Prothrombin Proteins 0.000 description 1
- 102100027378 Prothrombin Human genes 0.000 description 1
- 108091006197 SARS-CoV-2 Nucleocapsid Protein Proteins 0.000 description 1
- BUGBHKTXTAQXES-UHFFFAOYSA-N Selenium Chemical compound [Se] BUGBHKTXTAQXES-UHFFFAOYSA-N 0.000 description 1
- BQCADISMDOOEFD-UHFFFAOYSA-N Silver Chemical compound [Ag] BQCADISMDOOEFD-UHFFFAOYSA-N 0.000 description 1
- QJVKUMXDEUEQLH-UHFFFAOYSA-N [B].[Fe].[Nd] Chemical compound [B].[Fe].[Nd] QJVKUMXDEUEQLH-UHFFFAOYSA-N 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 239000000956 alloy Substances 0.000 description 1
- 229910045601 alloy Inorganic materials 0.000 description 1
- 229910000828 alnico Inorganic materials 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 244000052616 bacterial pathogen Species 0.000 description 1
- 108010005774 beta-Galactosidase Proteins 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- FCCCRBDJBTVFSJ-UHFFFAOYSA-N butanehydrazide Chemical compound CCCC(=O)NN FCCCRBDJBTVFSJ-UHFFFAOYSA-N 0.000 description 1
- WAWNVIAZVPDFOT-UHFFFAOYSA-N butanehydrazide;hydrochloride Chemical compound Cl.CCCC(=O)NN WAWNVIAZVPDFOT-UHFFFAOYSA-N 0.000 description 1
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 1
- 210000001175 cerebrospinal fluid Anatomy 0.000 description 1
- 230000021615 conjugation Effects 0.000 description 1
- 238000011033 desalting Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- LTMHDMANZUZIPE-PUGKRICDSA-N digoxin Chemical compound C1[C@H](O)[C@H](O)[C@@H](C)O[C@H]1O[C@@H]1[C@@H](C)O[C@@H](O[C@@H]2[C@H](O[C@@H](O[C@@H]3C[C@@H]4[C@]([C@@H]5[C@H]([C@]6(CC[C@@H]([C@@]6(C)[C@H](O)C5)C=5COC(=O)C=5)O)CC4)(C)CC3)C[C@@H]2O)C)C[C@@H]1O LTMHDMANZUZIPE-PUGKRICDSA-N 0.000 description 1
- 229960005156 digoxin Drugs 0.000 description 1
- LTMHDMANZUZIPE-UHFFFAOYSA-N digoxine Natural products C1C(O)C(O)C(C)OC1OC1C(C)OC(OC2C(OC(OC3CC4C(C5C(C6(CCC(C6(C)C(O)C5)C=5COC(=O)C=5)O)CC4)(C)CC3)CC2O)C)CC1O LTMHDMANZUZIPE-UHFFFAOYSA-N 0.000 description 1
- 239000000975 dye Substances 0.000 description 1
- 239000003792 electrolyte Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 1
- 238000009396 hybridization Methods 0.000 description 1
- GXBDUKNALWQPMX-UHFFFAOYSA-N hydron;pyrrole-2,5-dione;chloride Chemical compound Cl.O=C1NC(=O)C=C1 GXBDUKNALWQPMX-UHFFFAOYSA-N 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 239000000138 intercalating agent Substances 0.000 description 1
- 229910052742 iron Inorganic materials 0.000 description 1
- HWYHZTIRURJOHG-UHFFFAOYSA-N luminol Chemical compound O=C1NNC(=O)C2=C1C(N)=CC=C2 HWYHZTIRURJOHG-UHFFFAOYSA-N 0.000 description 1
- 239000002122 magnetic nanoparticle Substances 0.000 description 1
- 239000006249 magnetic particle Substances 0.000 description 1
- 238000000691 measurement method Methods 0.000 description 1
- 239000004005 microsphere Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 229910001172 neodymium magnet Inorganic materials 0.000 description 1
- 150000002843 nonmetals Chemical class 0.000 description 1
- 210000002381 plasma Anatomy 0.000 description 1
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 description 1
- 229920000136 polysorbate Polymers 0.000 description 1
- 229920000053 polysorbate 80 Polymers 0.000 description 1
- 229940039716 prothrombin Drugs 0.000 description 1
- 239000002096 quantum dot Substances 0.000 description 1
- 230000002285 radioactive effect Effects 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- SUKJFIGYRHOWBL-UHFFFAOYSA-N sodium hypochlorite Chemical compound [Na+].Cl[O-] SUKJFIGYRHOWBL-UHFFFAOYSA-N 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 229940124530 sulfonamide Drugs 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- LMYRWZFENFIFIT-UHFFFAOYSA-N toluene-4-sulfonamide Chemical compound CC1=CC=C(S(N)(=O)=O)C=C1 LMYRWZFENFIFIT-UHFFFAOYSA-N 0.000 description 1
- KAKQVSNHTBLJCH-UHFFFAOYSA-N trifluoromethanesulfonimidic acid Chemical compound NS(=O)(=O)C(F)(F)F KAKQVSNHTBLJCH-UHFFFAOYSA-N 0.000 description 1
- GPRLSGONYQIRFK-MNYXATJNSA-N triton Chemical compound [3H+] GPRLSGONYQIRFK-MNYXATJNSA-N 0.000 description 1
- 210000002700 urine Anatomy 0.000 description 1
- 229910000859 α-Fe Inorganic materials 0.000 description 1
Classifications
-
- 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
- G01N33/5306—Improving reaction conditions, e.g. reduction of non-specific binding, promotion of specific binding
-
- 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
- G01N33/531—Production of immunochemical test materials
-
- 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
- G01N33/531—Production of immunochemical test materials
- G01N33/532—Production of labelled immunochemicals
-
- 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
- G01N33/531—Production of immunochemical test materials
- G01N33/532—Production of labelled immunochemicals
- G01N33/533—Production of labelled immunochemicals with fluorescent label
-
- 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
- G01N33/531—Production of immunochemical test materials
- G01N33/532—Production of labelled immunochemicals
- G01N33/535—Production of labelled immunochemicals with enzyme label or co-enzymes, co-factors, enzyme inhibitors or enzyme substrates
-
- 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
- G01N33/536—Immunoassay; Biospecific binding assay; Materials therefor with immune complex formed in liquid phase
-
- 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
- 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
- G01N33/54313—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being characterised by its particulate form
- G01N33/54326—Magnetic particles
-
- 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
- G01N33/573—Immunoassay; Biospecific binding assay; Materials therefor for enzymes or isoenzymes
-
- 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
- G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
-
- 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
- G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
- G01N33/57473—Immunoassay; Biospecific binding assay; Materials therefor for cancer involving carcinoembryonic antigen, i.e. CEA
-
- 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
- G01N33/577—Immunoassay; Biospecific binding assay; Materials therefor involving monoclonal antibodies binding reaction mechanisms characterised by the use of monoclonal antibodies; monoclonal antibodies per se are classified with their corresponding antigens
-
- 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
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
-
- 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
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6863—Cytokines, i.e. immune system proteins modifying a biological response such as cell growth proliferation or differentiation, e.g. TNF, CNF, GM-CSF, lymphotoxin, MIF or their receptors
- G01N33/6869—Interleukin
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Immunology (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Molecular Biology (AREA)
- Urology & Nephrology (AREA)
- Biomedical Technology (AREA)
- Hematology (AREA)
- Cell Biology (AREA)
- Food Science & Technology (AREA)
- General Health & Medical Sciences (AREA)
- Biotechnology (AREA)
- Medicinal Chemistry (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- Microbiology (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Oncology (AREA)
- Hospice & Palliative Care (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Investigating Or Analyzing Materials By The Use Of Electric Means (AREA)
Abstract
本发明涉及一种稀释剂、捕获试剂、检测试剂盒和测定方法。该稀释剂包括缓冲剂和电导率调节剂,稀释剂的电导率在33mS/cm以上。该稀释剂可以降低检测的本底,提高检测的信噪比,提高检测的灵敏度。
Description
技术领域
本发明涉及生物检测技术领域,特别是涉及一种稀释剂、捕获试剂、检测试剂盒和测定方法。
背景技术
在免疫检测试剂盒研发过程中,为保证试剂盒质量,需按照指导文件对多项指标进行评估。灵敏度作为一项重要的技术指标,对试剂盒的评价具有关键意义,因为极少量的分析物可能对界定疾病状态、疾病筛查以及病情监测和治疗评估等具有重要指导意义。
发明内容
基于此,本发明提供一种能够提高免疫检测试剂盒的灵敏度的稀释剂。
此外,还提供一种上述稀释剂在制备免疫检测的捕获试剂中的应用、包括上述稀释剂的捕获试剂和检测试剂盒,以及应用上述稀释剂的样品中分析物的测定方法。
一种稀释剂,包括缓冲剂和电导率调节剂,所述稀释剂的工作电导率在33mS/cm以上,所述稀释剂用于处理包括磁珠的试剂。
电导率调节剂为能显著影响溶液电导率的电解质,经本申请发明人研究发现,在免疫检测过程中,将与待测样品中的分析物特异性结合的捕获试剂中的稀释剂的工作电导率设置在33mS/cm以上时,随着稀释剂的工作电导率的增大,检测到的信号值会呈下降趋势,可以提高检测的信噪比,进而提高检测的灵敏度。
在一实施例中,所述稀释剂的工作电导率在350mS/cm以下;
优选的,所述稀释剂的工作电导率为50mS/cm~320mS/cm。
在一实施例中,所述缓冲剂包括磷酸盐缓冲液、HEPES缓冲液、MOPS缓冲液、Tris-HCl缓冲液和MES缓冲液中的一种或多种。
在一实施例中,所述电导率调节剂包括盐;优选地,所述盐为钠盐、钾盐、铵盐、钙盐、镁盐中的至少一种。
在一实施例中,所述稀释剂还包括表面活性剂和防腐剂中的至少一种。
在一实施例中,所述盐为NaCl,在所述稀释剂中,所述NaCl的工作浓度为1%(m/v)~10%(m/v);
或者,所述盐为KCl,在所述稀释剂中,所述KCl的工作浓度为3%(m/v)~10%(m/v);
或者,所述盐为NH4Cl,在所述稀释剂中,所述NH4Cl的工作浓度为0.88%(m/v)~25%(m/v)。
上述的稀释剂在制备免疫检测的捕获试剂中的应用,所述捕获试剂能够与待测样品中的分析物特异性结合。
一种捕获试剂,所述捕获试剂能够与待测样品中的分析物特异性结合,所述捕获试剂包括上述的稀释剂,所述捕获试剂包括磁珠。
进一步地所述磁珠包被有能与所述分析物特异性结合的物质。
可选地,所述捕获试剂用于捕获分析物,可选地,所述分析物为抗体或抗原;
进一步地,所述分析物为CEA、NSE、PIVKA-II、SARS-CoV-2、SARS-CoV-2的N蛋白、SARS-CoV-2抗体、C-反应蛋白、血清淀粉样蛋白A、降钙素原、白细胞介素6、尿微量白蛋白、β2-微球蛋白、视黄醇结合蛋白、胱抑素C、人中性粒细胞明胶酶相关脂质运载蛋白、D-二聚体、肌钙蛋白I(例如心肌肌钙蛋白I)、肌钙蛋白T、N末端脑钠肽、N-末端脑钠肽前体、脑尿钠肽、肌酸激酶同工酶或肌红蛋白中的至少一种。
捕获试剂中含有磁珠,因此无法直接检测电导率,但可以过滤掉磁珠,再检测滤液电导率,滤液电导率应当基本与上述稀释剂电导率一致。
一种检测试剂盒,包括上述的捕获试剂。
进一步地,所述试剂盒还包括标记剂,所述标记剂包括连接有标记物且能与待测样品中的分析物特异性结合的物质。
进一步地,上述捕获试剂和标记剂涉及的能与分析物特异性结合的物质为抗体、抗原、配体、受体、寡核苷酸、半抗原、表位、模拟表位或适体中的至少一个。
一种样品中分析物的测定方法,包括以下步骤:
使待测样品、标记剂和捕获试剂接触,通过检测所述标记剂与所述待测样品中的分析物和所述捕获试剂形成的复合物的量而确定所述分析物的量,其中,所述捕获试剂为上述的捕获试剂。
具体实施方式
为了便于理解本发明,下面将对本发明进行更全面的描述。但是,本发明可以以许多不同的形式来实现,并不限于本文所描述的实施例。相反地,提供这些实施例的目的是使本发明公开内容更加透彻全面。
除非另有定义,本文所使用的所有的技术和科学术语与属于本发明的技术领域的技术人员通常理解的含义相同。本文中在本发明的说明书中所使用的术语只是为了描述具体的实施例的目的,不是旨在于限制本发明。术语“和/或”包括一个或多个相关的所列项目的任意的和所有的组合。本文中“mS/cm”是电导率的单位,也即毫西门子每厘米。本文中“可选地”用于举例。本文中术语“以上”在表示数值范围时,包括本数。需要说明的是,在本文中“稀释剂的工作电导率”是指稀释剂在使用时的电导率。
本申请一实施方式提供了一种稀释剂,该稀释剂包括缓冲剂和电导率调节剂,该稀释剂的工作电导率在33mS/cm以上,所述稀释剂用于处理包括磁珠的试剂。
经本申请发明人研究发现,在免疫检测过程中,将与待测样品中的分析物特异性结合的捕获试剂中的稀释剂的工作电导率设置在33mS/cm以上时,随着稀释剂的电导率的增大,检测到的信号值会呈下降趋势,可以降低检测的本底,提高检测的信噪比,提高检测的灵敏度。
在一些实施例中,稀释剂的工作电导率在350mS/cm以下。在一些可选地具体示例中,稀释剂的工作电导率为33mS/cm、45mS/cm、55mS/cm、70mS/cm、80mS/cm、100mS/cm、120mS/cm、150mS/cm、180mS/cm、200mS/cm、220mS/cm、250mS/cm、270mS/cm、300mS/cm、310mS/cm或350mS/cm。进一步地,稀释剂的工作电导率为50mS/cm~320mS/cm。
在一些实施例中,稀释剂的工作电导率为41.7mS/cm~85.78mS/cm。进一步地,稀释剂的工作电导率为65.1mS/cm~113.77mS/cm。
在另一些实施例中,稀释剂的工作电导率为65.46mS/cm~88.92mS/cm。
在另一些实施例中,稀释剂的工作电导率为50.75mS/cm~113.77mS/cm。进一步地,稀释剂的工作电导率为82.26mS/cm~113.77mS/cm。
在另一些实施例中,稀释剂的工作电导率为33.94mS/cm~320.87mS/cm。进一步地,稀释剂的工作电导率为50.75mS/cm~113.77mS/cm或50.75mS/cm~320.87mS/cm。
具体地,缓冲剂用于构建缓冲体系。可选地,缓冲剂含有磷酸盐缓冲液、HEPES缓冲液、MOPS缓冲液、Tris-HCl缓冲液、和MES缓冲液中的一种或多种。优选地,缓冲剂含有MES缓冲液。可以理解的是,缓冲剂不限于上述,还可以其他物质。
具体地,电导率调节剂用于调节稀释剂的电导率。可选地,电导率调节剂为能调节溶液离子强度的盐。在一些实施例中,盐包括NaCl、KCl和NH4Cl、MgCl2、MgSO4、CaCl2、Na2SO4、K2SO4、NH4SO4、NaNO3、KNO3、KClO4、NaClO4中的至少一种。优选地,盐包括NaCl、KCl和NH4Cl中的至少一种。
在一些实施例中,盐为NaCl,在稀释剂中NaCl的工作浓度为1.8%(m/v)~10%(m/v)。进一步地,在稀释剂中,在含有0.15mol/L NaCl的基础上添加NaCl,NaCl的添加量为1.5%(m/v)~8%(m/v)。更进一步地,在稀释剂中NaCl的工作浓度为1.75%(m/v)~5.5%(m/v)。需要说明的是,m/v表示质量体积浓度,例如,10%(m/v)是指100mL中含有10g溶质,其他类推。本文中,NaCl的工作浓度是指在使用时稀释剂中NaCl的浓度。
在一些实施例中,盐为NaCl,稀释剂的工作电导率为33mS/cm~100mS/cm。进一步地,稀释剂的工作电导率为41.7mS/cm~85.78mS/cm。更进一步地,稀释剂的工作电导率为65.1mS/cm~85.78mS/cm。
在一些实施例中,盐为NaCl,在稀释剂中NaCl的工作浓度为1%~10%(m/v);稀释剂的工作电导率为33mS/cm~100mS/cm。进一步地,在稀释剂中NaCl的工作浓度为1.75%(m/v)~5.26%(m/v);稀释剂的工作电导率为41.7mS/cm~85.78mS/cm。更进一步地,在稀释剂中NaCl的工作浓度为3.51%(m/v)~5.26%(m/v),稀释剂的工作电导率为65.1mS/cm~85.78mS/cm。
在一些实施例中,盐为KCl,在稀释剂中,KCl的工作浓度为3%(m/v)~10%(m/v)。进一步地,在稀释剂中,KCl的工作浓度为3.5%(m/v)~6%(m/v)。本文中,KCl的工作浓度是指在使用时稀释剂中KCl的浓度。
在一些实施例中,盐为KCl,稀释剂的工作电导率为45mS/cm~100mS/cm。进一步地,稀释剂的工作电导率为65.46mS/cm~88.92mS/cm。
在一些实施例中,盐为KCl,在稀释剂中,KCl的工作浓度为3%(m/v)~10%(m/v);稀释剂的工作电导率为45mS/cm~100mS/cm。进一步地,在稀释剂中,KCl的工作浓度为3.51%(m/v)~5.26%(m/v);稀释剂的工作电导率为65.46mS/cm~88.92mS/cm。
在一些实施例中,盐为NH4Cl,在稀释剂中,NH4Cl的工作浓度为0.88%(m/v)~25%(m/v)。进一步地,在稀释剂中,NH4Cl的工作浓度为1.5%(m/v)~22%(m/v)。本文中,NH4Cl的工作浓度是指在使用时稀释剂中NH4Cl的浓度。
在一些实施例中,盐为NH4Cl,稀释剂的工作电导率为33.94mS/cm~340mS/cm。进一步地,稀释剂的工作电导率为40mS/cm~320mS/cm。
在一些实施例中,盐为NH4Cl,在稀释剂中,NH4Cl的工作浓度为0.88%(m/v)~25%(m/v);稀释剂的工作电导率为33.94mS/cm~340mS/cm。进一步地,在稀释剂中,NH4Cl的工作浓度为1.5%(m/v)~22%(m/v)。稀释剂的工作电导率为40mS/cm~320mS/cm。
在其中一个实施例中,盐为NH4Cl,在稀释剂中NH4Cl的工作浓度为1%(m/v)~10%(m/v);稀释剂的工作电导率为40mS/cm~120mS/cm。进一步地,在稀释剂中,NH4Cl的工作浓度为1.5%(m/v)~8%(m/v);稀释剂的工作电导率为40mS/cm~120mS/cm。更进一步地,在稀释剂中,NH4Cl的工作浓度为1.75%(m/v)~5.26%(m/v);稀释剂的工作电导率为50.75mS/cm~113.77mS/cm。
在其中一个实施例中,盐为NH4Cl,在稀释剂中NH4Cl的工作浓度为0.88%(m/v)~21.04%(m/v);稀释剂的工作电导率为33.94mS/cm~320.87mS/cm。进一步地,在稀释剂中,NH4Cl的工作浓度为1.5%(m/v)~21%(m/v)。稀释剂的工作电导率为50mS/cm~320mS/cm。
在一些实施例中,上述稀释剂还包括表面活性剂和防腐剂中的至少一种。可选地,表面活性剂为Tween20、Tween80和Tween100中的至少一种。可选地,防腐剂为Proclin300。可以理解的是,表面活性剂和防腐剂均不限于上述,还可以是其他物质。
基于上述,本申请一实施方式还提供了一种上述任一实施例的稀释剂在制备检测产品中的应用。可选地,检测产品为检测试剂盒或检测试剂(例如捕获试剂)。
具体地,上述任一实施例的稀释剂在制备免疫检测的捕获试剂中的应用,其中,捕获试剂能够与待测样品中的分析物特异性结合。
此外,本申请一实施方式还提供了一种检测试剂盒,该检测试剂盒包括捕获试剂和标记剂,捕获试剂包括上述任一实施例的稀释剂,标记剂上连接有标记物,标记剂能与待测样品中的分析物特异性结合。
具体地,捕获试剂基于免疫检测技术,能够与待测样品中的分析物特异性结合,用于捕获分析物;进一步地,捕获试剂包括磁珠和上述任一实施例的稀释剂;捕获试剂中的磁珠能与待测样品中的分析物特异性结合而捕获分析物。连接有标记物的标记剂起标记分析物的作用,用于反馈分析物的量。分析物是同时能与捕获试剂和标记剂特异性结合的物质。分析物与捕获试剂和标记剂以结合对的形式特性结合。例如受体/配体对、抗体/抗原、天然或合成受体/配体对、半抗原/抗体对、抗原/抗体对、表位/抗体对、模拟位/抗体对、适体/靶分子对、杂交伴侣、和嵌入物/靶分子对。当然,在基于夹心法的免疫检测中,捕获试剂与分析物的结合位点与标记剂和分析物的结合位点不同。在基于竞争法的免疫检测中,捕获试剂与分析物的结合位点与标记剂和分析物的结合位点相同。可选地,分析物为癌胚抗原(CEA)、神经元特异性烯醇化酶(NSE)、异常凝血酶原(DCP,又称PIVKA-II)、SARS-CoV-2、SARS-CoV-2的N蛋白、SARS-CoV-2抗体、C-反应蛋白、血清淀粉样蛋白A、降钙素原、白细胞介素6、尿微量白蛋白、β2-微球蛋白、视黄醇结合蛋白、胱抑素C、人中性粒细胞明胶酶相关脂质运载蛋白、D-二聚体、肌钙蛋白I(例如心肌肌钙蛋白I)、肌钙蛋白T、N末端脑钠肽、N-末端脑钠肽前体、脑尿钠肽、肌酸激酶同工酶或肌红蛋白(MYO)中的至少一种。
在一些实施例中,磁珠,其成分中含有磁性物质。在一些实施例中,磁珠的直径为10nm~1mm。在一个可选地具体示例中,磁珠的直径为100nm、500nm、1μm、10μm、100μm或500μm。进一步地,磁珠的直径为400nm~10μm。磁性物质可以为金属(金属单质或合金)、非金属,或金属与非金属所形成的复合物。金属例如铁、铝镍钴金属等;非金属例如铁氧体非金属(例如Fe2O3或Fe3O4磁性纳米粒子);金属与非金属所形成的复合物例如钕铁硼橡胶磁复合材料。在一些实施例中,磁珠的表面修饰有一种或多种活性功能基团。可选地,活性功能基团包括-OH、-COOH、-NH2、-CHO及-SO3H中的一种或多种。在一些实施例中,包被的连接部通过物理吸附或直接化学缀合(例如通过桥接物进行桥接)与磁珠缀合或结合。在一个可选地具体示例中,磁珠通过链霉亲和素和生物素连接有抗体。在另一个可选地具体示例中,磁珠上连接有链霉亲和素。
本文中,标记剂上连接的标记物是指能够提供被检测的信号的物质。在一些实施例中,标记物选自发色团、地高辛标记探针、电子致密物质、金属粒子及产生可检测信号的酶中的至少一种。可选地,发色团选自荧光、量子点、荧光微球、发光化合物和染料中的一种或多种。在一些实施例中,发色团为发光化合物。例如,吖啶酯、吖啶酯衍生物、金刚烷、鲁米诺、异鲁米诺等。可选地,吖啶酯衍生物选自吖啶酯磺酰胺、吖啶酯甲苯磺酰胺、吖啶酯对甲基磺酰胺及吖啶酯三氟甲基磺酰胺中的至少一种。电子致密物质为放射性分子。例如32P,35S或125I。在一些实施例中,产生可检测信号的酶选自辣根过氧化物酶、碱性磷酸酶、β-半乳糖苷酶和葡萄糖-6-磷酸脱氢酶中的一种。在一些实施例中,金属粒子是胶体金属,例如胶体金、胶体银、胶体硒等。可以理解的是,标记物不限于上述,还可以是其他能够被肉眼直接观察到、借助仪器检测到的物质。
在一些实施例中,标记物直接与标记剂连接。
在另一些实施例中,标记物通过桥接物间接与标记剂连接。可选地,桥接物选自蛋白、蛋白复合物或双功能交联剂中的一种或多种。在其中一个实施例中,作为桥接物的蛋白中含有牛血清白蛋白、卵白蛋白、钥孔血蓝蛋白、免疫球蛋白、甲状腺球蛋白或多聚赖氨酸。作为桥接物的蛋白复合物中含有牛血清白蛋白、卵白蛋白、钥孔血蓝蛋白、免疫球蛋白、甲状腺球蛋白和多聚赖氨酸中的至少两种。可选地,双功能交联剂选自4-(4-N-马来酰亚胺基苯基)丁酸酰肼(4-[4-N-maleimidophenyl]butyric acid hydrazide hydrochloride,MPBH)、1-[2-[2-(2-氨基乙氧基)乙氧基]乙基]马来酰亚胺盐酸盐(1-[2-[2-(2-Aminoethoxy)ethoxy]ethyl]maleimide hydrochloride,MPEG2A)、N-[β-马来酰亚胺丙酸]酰肼,三氟乙酸盐(N-[β-maleimidopropionic acid]hydrazide,trifluoroacetic acidsalt,BMPH)、N-[ε-马来酰亚胺基己酸)酰肼,三氟乙酸盐(N-[ε-Maleimidocaproic acid)hydrazide,trifluoroacetic acid salt,EMCH)、N-[κ-马来酰亚胺十一烷酸]酰肼,三氟乙酸盐(N-[κ-maleimidoundecanoic acid]hydrazide,trifluoroacetic acid salt,KMUH)中的任一种。可以理解的是,在一些实施例中,桥接物具有放大信号的作用。例如,桥接物可以偶联更多的标记物以放大信号。
可选地,待测样品是尿液、脑脊髓液、粪便、血液、血清或血浆。可以理解的是,待测样品不限于上述。
可以理解的是,在一些实施例中,上述检测试剂盒中的标记剂可以省略。标记剂省略时,上述检测试剂盒与标记剂(例如另购)搭配使用即可。
上述捕获试剂或上述检测试剂盒包括上述稀释剂,具有上述稀释剂相应的优点。
基于上述,本申请一实施方式还提供了一种样品中分析物的测定方法,包括以下步骤:将待测样品、标记剂和上述任一实施例的捕获试剂混合反应后,通过检测标记剂与待测样品中的分析物和捕获试剂形成的复合物的量而确定分析物的量。
在一些实施例中,该测定方法不以疾病的诊断或治疗为直接目的。例如,该测定方法用于海关检疫、用于公共场所反应卫生状况中(例如条件致病菌)的检测等。当然,在其他实施例中,该测定方法还可以辅助疾病的诊断或治疗。
进一步地,该测定方法基于夹心法进行测定。在混合反应后,分析物与标记剂和捕获剂中的磁珠共同形成夹心复合物,通过检测夹心复合物的量可以确定待测样品中分析物的量。
进一步地,该测定方法基于竞争法进行测定。在混合反应时,分析物与标记剂竞争性结合磁珠,所以通过检测标记剂和磁珠形成的复合物的量可以确定待测样品中分析物的量。
上述测定方法采用上述稀释剂作为捕获试剂的稀释剂,灵敏度高。
具体实施例
以下结合具体实施例进行详细说明。以下实施例如未特殊说明,则不包括除不可避免的杂质外的其他组分。实施例中采用试剂和仪器如非特别说明,均为本领域常规选择。实施例中未注明具体条件的实验方法,按照常规条件,例如文献、书本中所述的条件或者生产厂家推荐的方法实现。
实施例1
为探究磁珠稀释液的电导率对NSE检测试剂灵敏度的影响,对多种电导率不同的磁珠稀释液进行研究。
(1)磁珠包被物浓缩液的制备:
(a)磁珠包被管中添加磁珠悬浮液,将包被管置于磁分离架上静置,待悬浮液澄清后移去上清液;向包被管中加入0.05mol/L pH6.0MES缓冲液,涡旋混合后,磁分离;
(b)再次向包被管中加入MES缓冲液,涡旋混匀,然后加入10mg/mL EDC溶液,终浓度1mg/mL,涡旋混匀,25℃反应半个小时;
(c)磁分离,加入一定量的MES缓冲液和NSE单克隆抗体1,包被量为20μg/mg,磁珠浓度10mg/mL,涡旋混匀;
(d)将包被管置于25度恒温摇床上,保持旋转混合反应2小时,磁珠与抗体发生共价偶联反应;
(e)将包被管置于磁分离架上磁分离,加入封闭液(Tris缓冲液,含BSA,Tween20,Proclin300)涡旋混匀,除去未结合的抗体;
(f)将包被管置于磁分离架上磁分离,加入封闭液,将包被管置于37度恒温摇床上,保持旋转混合反应16小时,封闭磁珠包被物;
(g)将包被管置于磁分离架上磁分离,加入磁珠保存液(Tris缓冲液,含BSA,Tween20,Proclin300)涡旋混匀,洗涤;
(h)将包被管置于磁分离架上磁分离,加入磁珠保存液,制得磁珠包被物浓缩液,置于2~8度保存。
(2)吖啶酯标记物浓缩液的制备:
将NSE单克隆抗体2溶液进行脱盐处理,置换至PB 7.2缓冲溶液中。向含有NSE单克隆抗体2的缓冲溶液中加入吖啶酯溶液进行交联,吖啶酯:抗体=20:1(摩尔比),震荡混匀,室温避光,静置反应60分钟。随后,向上述交联溶液中加入甘氨酸溶液,甘氨酸:吖啶酯=20:1(摩尔比),迅速震荡混匀,室温避光,静置反应30分钟。将交联封闭完成的溶液进行脱盐,并置换至PBS 7.4缓冲溶液中,加入甘油,-20℃保存。
(3)各组份工作液的配制:
配制原磁珠包被物稀释液(成分包括:25mM MES、0.15mol/L NaCl、1.0%(m/v)酪蛋白钠盐、0.05%(v/v)Tween-20、0.05%(v/v)Tritonx-100),以及在此基础上分别添加1.75%、3.51%和5.26%的NaCl或KCl或NH4Cl(1.75%NaCl是指按照100mL原磁珠包被物稀释液1.75g的NaCl的比例加入NaCl,5.26%KCl指按照100mL原磁珠包被物稀释液5.26g的KCl的比例加入KCl,其他依次类推),从而制得8种电导率不同的溶液。分别使用上述8种磁珠包被物稀释液按同等比例稀释步骤(1)制备的磁珠包被物浓缩液,混匀后即制得8种磁珠组份工作液,磁珠组份工作液中磁珠的含量为0.5mg/mL。
使用吖啶酯标记物稀释液(组成为:25mM PB、0.03mol/L NaCl、1.0%(m/v)酪蛋白钠盐/0.1%(v/v)Tween-20以及多少Proclin300)按一定比例稀释吖啶酯标记物浓缩液,混匀后即制得吖啶酯组份工作液,吖啶酯组份工作液中吖啶标记物的浓度为:0.5μg/mL。其中,吖啶酯标记物稀释液的电导率的均值为5.938mS/cm(三次测定的值分别为5.873mS/cm、5.958mS/cm、5.984mS/cm)。
(4)检测方式
使用Shine i2000全自动化学发光免疫分析仪检测不同条件下NSE零值和高值样本的信号值。反应程序为:依次加入10μL样本、50μL磁珠组份工作液和50μL吖啶酯组份工作液,混匀后于37℃反应15min,使用洗液TBST清洗4次,加入激发液A(含H2O2的酸性溶液)和B(含NaOH的溶液),仪器读数。
结果如表1所示。在表1中,原磁珠稀释液是指原磁珠包被物稀释液,具体组成如上述。
表1
采用化学发光试剂检测NSE零值和高值样本,对多种电导率不同的磁珠包被物稀释液进行研究。由表1可知,随着磁珠包被物稀释液电导率的增大,高值和零值样本的信号值均存在不同幅度下降的现象,并且在一定范围内,试剂检测灵敏度提高。具体来说,本研究中使用的3种不同盐离子均呈现出同样的趋势,即当磁珠稀释液的电导率在41.70~113.77mS/cm时,试剂检测信噪比提高1.6~3.0倍。
实施例2
为探究磁珠包被物稀释液的电导率对PIVKA-II检测试剂灵敏度的影响,对多种电导率不同的磁珠包被物稀释液进行研究。
(1)磁珠包被物浓缩液的制备:
(a)磁珠包被管中添加磁珠悬浮液,将包被管置于磁分离架上静置,待悬浮液澄清后移去上清液;向包被管中加入0.05mol/L pH6.0 MES缓冲液,涡旋混合后,磁分离;
(b)再次向包被管中加入MES缓冲液,涡旋混匀,然后加入10mg/mL EDC溶液,终浓度1mg/mL,涡旋混匀,25℃反应半个小时;
(c)磁分离,加入一定量的MES缓冲液和亲和素(SA),包被量为20μg/mg,磁珠浓度10mg/mL,涡旋混匀;
(d)将包被管置于25度恒温摇床上,保持旋转混合反应2小时,磁珠与亲和素发生共价偶联反应;
(e)将包被管置于磁分离架上磁分离,加入封闭液(Tris缓冲液,含BSA,Tween20,Proclin300)涡旋混匀,除去未结合的抗体;
(f)将包被管置于磁分离架上磁分离,加入封闭液,将包被管置于37度恒温摇床上,保持旋转混合反应16小时,封闭磁珠包被物;
(g)将包被管置于磁分离架上磁分离,加入磁珠保存液(Tris缓冲液,含BSA,Tween20,Proclin300)涡旋混匀,洗涤;
(h)将包被管置于磁分离架上磁分离,加入磁珠保存液,制得磁珠包被物浓缩液,置于2~8度保存。
(2)生物素标记物浓缩液的制备:
将PIVKA-II单克隆抗体1进行脱盐处理,置换至PB7.2缓冲溶液中。向含PIVKA-II单克隆抗体1的缓冲溶液中加入生物素溶液,生物素:抗体=30:1(摩尔比),震荡混匀,室温避光,静置反应60分钟。随后,向上述反应后的溶液中加入甘氨酸溶液,甘氨酸:生物素=30:1(摩尔比),迅速震荡混匀,室温避光,静置反应30分钟。将交联封闭完成的溶液进行脱盐,并置换至PBS 7.4缓冲溶液中,加入甘油,-20℃保存。
(3)吖啶酯标记物浓缩液的制备:
将PIVKA-II单克隆抗体2溶液进行脱盐处理,置换至PB 7.2缓冲溶液中。向含有PIVKA-II单克隆抗体2的缓冲溶液中加入吖啶酯溶液进行交联,吖啶酯:抗体=20:1(摩尔比),震荡混匀,室温避光,静置反应60分钟。随后,向上述交联溶液中加入甘氨酸溶液,甘氨酸:吖啶酯=20:1(摩尔比),迅速震荡混匀,室温避光,静置反应30分钟。将交联封闭完成的溶液进行脱盐,并置换至PBS7.4缓冲溶液中,加入甘油,-20℃保存。
(4)各组份工作液的配制:
(a)配制原磁珠包被物稀释液(组成同实施例1),以及在此基础上分别添加0.88%、1.75%、3.51%、10.52%和21.04%的NH4Cl(0.88%NH4Cl是指按照100mL原磁珠包被物稀释液0.88g的NH4Cl的比例加入NH4Cl,其他同理),从而制得5种电导率不同的溶液。分别使用上述5种磁珠稀释液按同等比例稀释磁珠包被物浓缩液,混匀后即制得5种磁珠组份工作液,磁珠组份工作液中磁珠的含量为0.5mg/mL。
(b)使用生物素标记物稀释液(组成同实施例1的原磁珠包被物稀释液)按一定比例稀释步骤(2)制得的生物素标记物浓缩液混匀后即制得生物素组份工作液,生物素组份工作液中生物素的含量为0.5μg/mL。
(c)使用吖啶酯标记物稀释液(同实施例1)按一定比例稀释吖啶酯标记物浓缩液,混匀后即制得吖啶酯组份工作液,吖啶酯组份工作液中吖啶酯的工作浓度为0.5μg/mL。吖啶酯标记物稀释液的电导率的均值为5.938mS/cm(三次测定的值分别为5.837mS/cm、5.958mS/cm、5.984mS/cm)。
(5)检测方式
使用Shine i2000全自动化学发光免疫分析仪检测不同条件下PIVKA-II零值和高值样本的信号值。反应程序为:依次加入30μL样本、50μL生物素组份工作液和50μL吖啶酯组份工作液,混匀后于37℃反应10min,加入50μL链霉亲和素磁性粒子组份工作液,混匀后于37℃反应10min,使用洗液TBST清洗4次,加入激发液A(含H2O2的酸性溶液)和B(含NaOH的溶液),仪器读数。
结果如表2所示。在表2中,原磁珠稀释液是指同实施例1。
表2
由表2可知。对于PIVKA-II检测试剂而言,在一定范围内增加磁珠包被物稀释液的电导率(33.94~320.87mS/cm),同样存在检测灵敏度提高的现象。在所测试的条件中,当电导率为50.75mS/cm时,试剂检测信噪比提高了2.74倍。
以上所述实施例的各技术特征可以进行任意的组合,为使描述简洁,未对上述实施例中的各个技术特征所有可能的组合都进行描述,然而,只要这些技术特征的组合不存在矛盾,都应当认为是本说明书记载的范围。
以上所述实施例仅表达了本发明的几种实施方式,便于具体和详细地理解本发明的技术方案,但并不能因此而理解为对发明专利保护范围的限制。应当指出的是,对于本领域的普通技术人员来说,在不脱离本发明构思的前提下,还可以做出若干变形和改进,这些都属于本发明的保护范围。应当理解的是,在本领域技术人员在本发明提供的技术方案的基础上,通过合乎逻辑的分析、推理或有限的试验得到的技术方案,均在本发明所附权利要求的保护范围内。因此,本发明专利的保护范围应以所附权利要求的内容为准,说明书可以用于解释权利要求的内容。
Claims (10)
1.一种稀释剂,其特征在于,包括缓冲剂和电导率调节剂,所述稀释剂的工作电导率在33mS/cm以上,所述稀释剂用于处理包括磁珠的试剂。
2.根据权利要求1所述的稀释剂,其特征在于,所述稀释剂的工作电导率在350mS/cm以下;
优选的,所述稀释剂的工作电导率为50mS/cm~320mS/cm。
3.根据权利要求1~2任一项所述的稀释剂,其特征在于,所述缓冲剂包括磷酸盐缓冲液、HEPES缓冲液、MOPS缓冲液、Tris-HCl缓冲液和MES缓冲液中的一种或多种;
和/或,所述电导率调节剂包括盐。
4.根据权利要求3所述的稀释剂,其特征在于,所述盐为钠盐、钾盐、铵盐、钙盐、镁盐中的至少一种。
5.权利要求1~4任一项所述的稀释剂在制备免疫检测的捕获试剂中的应用,所述捕获试剂能够与待测样品中的分析物特异性结合。
6.一种捕获试剂,其特征在于,所述捕获试剂能够与待测样品中的分析物特异性结合,所述捕获试剂包括权利要求1~5任一项所述的稀释剂,所述捕获试剂包括磁珠。
7.根据权利要求6所述的捕获试剂,其特征在于,所述分析物为抗体或抗原;
进一步地,所述分析物为CEA、NSE、PIVKA-II、SARS-CoV-2、SARS-CoV-2的N蛋白、SARS-CoV-2抗体、C-反应蛋白、血清淀粉样蛋白A、降钙素原、白细胞介素6、尿微量白蛋白、β2-微球蛋白、视黄醇结合蛋白、胱抑素C、人中性粒细胞明胶酶相关脂质运载蛋白、D-二聚体、肌钙蛋白I(例如心肌肌钙蛋白I)、肌钙蛋白T、N末端脑钠肽、N-末端脑钠肽前体、脑尿钠肽、肌酸激酶同工酶或肌红蛋白中的至少一种。
8.根据权利要求6所述的捕获试剂,其特征在于,所述磁珠包被有能与所述分析物特异性结合的物质。
9.一种检测试剂盒,其特征在于,包括权利要求6或7所述的捕获试剂,
任选地,所述试剂盒还包括标记剂,所述标记剂包括连接有标记物且能与待测样品中的分析物特异性结合的物质。
10.一种样品中分析物的测定方法,其特征在于,包括以下步骤:
使待测样品、标记剂和捕获试剂接触,通过检测所述标记剂与所述待测样品中的分析物和所述捕获试剂形成的复合物的量而确定所述分析物的量,其中,所述捕获试剂为权利要求6、7或8所述的捕获试剂。
Priority Applications (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202210734085.0A CN117347616A (zh) | 2022-06-27 | 2022-06-27 | 稀释剂、捕获试剂、检测试剂盒和测定方法 |
PCT/CN2023/102559 WO2024002038A1 (zh) | 2022-06-27 | 2023-06-27 | 稀释剂、捕获试剂、检测试剂盒和测定方法 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202210734085.0A CN117347616A (zh) | 2022-06-27 | 2022-06-27 | 稀释剂、捕获试剂、检测试剂盒和测定方法 |
Publications (1)
Publication Number | Publication Date |
---|---|
CN117347616A true CN117347616A (zh) | 2024-01-05 |
Family
ID=89363640
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202210734085.0A Pending CN117347616A (zh) | 2022-06-27 | 2022-06-27 | 稀释剂、捕获试剂、检测试剂盒和测定方法 |
Country Status (2)
Country | Link |
---|---|
CN (1) | CN117347616A (zh) |
WO (1) | WO2024002038A1 (zh) |
Family Cites Families (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5753094A (en) * | 1995-09-20 | 1998-05-19 | Beckman Instruments, Inc. | Borate storage buffer and sample diluent |
WO2021039492A1 (ja) * | 2019-08-30 | 2021-03-04 | コニカミノルタ株式会社 | 検体希釈液、標識化抗体分散液、サンドイッチ法 |
CN112881675A (zh) * | 2019-11-29 | 2021-06-01 | 广东菲鹏生物有限公司 | IgM抗体检测稀释液 |
CN114371288A (zh) * | 2020-10-15 | 2022-04-19 | 广东菲鹏生物有限公司 | 一种标记稀释液 |
CN114252595A (zh) * | 2021-12-23 | 2022-03-29 | 武汉生之源生物科技股份有限公司 | 一种降低样本基质干扰的磁珠稀释液及免疫检测试剂盒 |
-
2022
- 2022-06-27 CN CN202210734085.0A patent/CN117347616A/zh active Pending
-
2023
- 2023-06-27 WO PCT/CN2023/102559 patent/WO2024002038A1/zh unknown
Also Published As
Publication number | Publication date |
---|---|
WO2024002038A1 (zh) | 2024-01-04 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN108700584B (zh) | 标记复合物及其制备方法、试剂盒、应用和检测系统 | |
Zhang et al. | Aptamer-based fluorometric lateral flow assay for creatine kinase MB | |
JP5200003B2 (ja) | 磁場を用いた試料中の標的分子の検出 | |
JP6014091B2 (ja) | 検体中の測定対象成分の測定方法及び測定用キット | |
US7018849B2 (en) | Process for (A) separating biological/ligands from dilute solutions and (B) conducting an immunochromatographic assay thereof employing superparamagnetic particles throughtout | |
WO2018047793A1 (ja) | 腫瘍マーカーの測定方法及び測定試薬 | |
EP3315969B1 (en) | Method of detecting test substance by immune complex transfer method | |
CN108802360B (zh) | 一种血清中可交换铜和铜蓝蛋白一步同时检测用试剂盒、制备方法及应用 | |
WO2019088142A1 (ja) | バイオアッセイのための検出剤及びそれを用いたシグナルの増幅方法 | |
Tao et al. | An ultrasensitive chemiluminescence immunoassay of chloramphenicol based on gold nanoparticles and magnetic beads | |
US4640898A (en) | Homogeneous fluorescence ligang binding assay based upon preferential alteration of the respective intensities of bound and free label by solvent components | |
JP3961559B2 (ja) | ブロック化酵素プローブ複合体 | |
EP3511712B1 (en) | Method for measuring thyroglobulin | |
EP2815238A2 (en) | A process for detection and optional quantification of an analyte | |
JP2022152733A (ja) | 抗体結合磁性粒子の保存安定化方法 | |
CN117347616A (zh) | 稀释剂、捕获试剂、检测试剂盒和测定方法 | |
KR20200135110A (ko) | 스위치 성 부착반응을 이용한 친화 분리 시스템 및 방법 | |
Khramtsov et al. | Nuclear magnetic resonance immunoassay of tetanus antibodies based on the displacement of magnetic nanoparticles | |
Ivanova et al. | Magnetic nanoparticle‐based fluorescent immunoassay for determination of progesterone in milk | |
CN117347617A (zh) | 稀释剂及其应用、样品中分析物的测定方法 | |
WO2024090399A1 (ja) | 検体中の被験物質の検出方法 | |
EP1597579A2 (en) | Generic method for latex agglutination assays | |
Khramtsov et al. | Development of an Immunosorbent for Solid-Phase NMR-Based Assay | |
EP3978925A1 (en) | Affinity separation system and method using switch-like adhesion reaction | |
CN113341131A (zh) | 用于检测伏马毒素b1的试剂盒及检测方法 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination |