CN116068191A - 一种含有icam1蛋白片段的脑梗塞生物标记物及其应用 - Google Patents
一种含有icam1蛋白片段的脑梗塞生物标记物及其应用 Download PDFInfo
- Publication number
- CN116068191A CN116068191A CN202210918772.8A CN202210918772A CN116068191A CN 116068191 A CN116068191 A CN 116068191A CN 202210918772 A CN202210918772 A CN 202210918772A CN 116068191 A CN116068191 A CN 116068191A
- Authority
- CN
- China
- Prior art keywords
- cerebral infarction
- antigen
- protein
- protein fragment
- samples
- 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
- 239000012634 fragment Substances 0.000 title claims abstract description 73
- 206010008118 cerebral infarction Diseases 0.000 title claims abstract description 61
- 208000026106 cerebrovascular disease Diseases 0.000 title claims abstract description 60
- 101000599852 Homo sapiens Intercellular adhesion molecule 1 Proteins 0.000 title claims abstract description 22
- 102100037877 Intercellular adhesion molecule 1 Human genes 0.000 title claims abstract description 22
- 239000000090 biomarker Substances 0.000 title claims abstract description 12
- 108091007433 antigens Proteins 0.000 claims abstract description 80
- 102000036639 antigens Human genes 0.000 claims abstract description 80
- 238000012360 testing method Methods 0.000 claims abstract description 7
- 102000004219 Brain-derived neurotrophic factor Human genes 0.000 claims description 11
- 108090000715 Brain-derived neurotrophic factor Proteins 0.000 claims description 11
- 239000003153 chemical reaction reagent Substances 0.000 claims description 8
- 239000000427 antigen Substances 0.000 abstract description 60
- 238000001514 detection method Methods 0.000 abstract description 28
- 239000012472 biological sample Substances 0.000 abstract description 4
- 108090000623 proteins and genes Proteins 0.000 description 47
- 102000004169 proteins and genes Human genes 0.000 description 42
- 230000035945 sensitivity Effects 0.000 description 32
- 210000002966 serum Anatomy 0.000 description 16
- 239000000523 sample Substances 0.000 description 15
- 206010008111 Cerebral haemorrhage Diseases 0.000 description 11
- 101000849321 Homo sapiens Protein RRP5 homolog Proteins 0.000 description 11
- 102100033976 Protein RRP5 homolog Human genes 0.000 description 11
- 102000000380 Matrix Metalloproteinase 1 Human genes 0.000 description 10
- 210000004027 cell Anatomy 0.000 description 10
- 208000010125 myocardial infarction Diseases 0.000 description 10
- 101001013150 Homo sapiens Interstitial collagenase Proteins 0.000 description 9
- 101001042393 Homo sapiens LIM and senescent cell antigen-like-containing domain protein 4 Proteins 0.000 description 8
- 102100021812 LIM and senescent cell antigen-like-containing domain protein 4 Human genes 0.000 description 8
- 238000000034 method Methods 0.000 description 8
- 239000007788 liquid Substances 0.000 description 7
- 238000012216 screening Methods 0.000 description 7
- 108010084867 N-methyl D-aspartate receptor subtype 2A Proteins 0.000 description 6
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 6
- 101001082131 Homo sapiens Pumilio homolog 3 Proteins 0.000 description 5
- 102100027358 Pumilio homolog 3 Human genes 0.000 description 5
- 239000000243 solution Substances 0.000 description 5
- 238000005406 washing Methods 0.000 description 5
- 102100022630 Glutamate receptor ionotropic, NMDA 2B Human genes 0.000 description 4
- 101710195187 Glutamate receptor ionotropic, NMDA 2B Proteins 0.000 description 4
- 239000011248 coating agent Substances 0.000 description 4
- 238000000576 coating method Methods 0.000 description 4
- 201000010099 disease Diseases 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 238000003556 assay Methods 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 238000003745 diagnosis Methods 0.000 description 3
- 238000010790 dilution Methods 0.000 description 3
- 239000012895 dilution Substances 0.000 description 3
- 238000000605 extraction Methods 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 238000000746 purification Methods 0.000 description 3
- 238000007789 sealing Methods 0.000 description 3
- 208000024891 symptom Diseases 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- 102100040124 Apoptosis-inducing factor 1, mitochondrial Human genes 0.000 description 2
- 102100030802 Beta-2-glycoprotein 1 Human genes 0.000 description 2
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 2
- 102100033933 Endoplasmic reticulum protein SC65 Human genes 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 102100026693 FAS-associated death domain protein Human genes 0.000 description 2
- 102100029458 Glutamate receptor ionotropic, NMDA 2A Human genes 0.000 description 2
- 101000890622 Homo sapiens Apoptosis-inducing factor 1, mitochondrial Proteins 0.000 description 2
- 101000793425 Homo sapiens Beta-2-glycoprotein 1 Proteins 0.000 description 2
- 101000639962 Homo sapiens Endoplasmic reticulum protein SC65 Proteins 0.000 description 2
- 101000911074 Homo sapiens FAS-associated death domain protein Proteins 0.000 description 2
- 101000713613 Homo sapiens Tubulin beta-4B chain Proteins 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- 208000032382 Ischaemic stroke Diseases 0.000 description 2
- 101710135898 Myc proto-oncogene protein Proteins 0.000 description 2
- 102100038895 Myc proto-oncogene protein Human genes 0.000 description 2
- 206010028851 Necrosis Diseases 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 101710150448 Transcriptional regulator Myc Proteins 0.000 description 2
- 102100036821 Tubulin beta-4B chain Human genes 0.000 description 2
- 230000036770 blood supply Effects 0.000 description 2
- 229940098773 bovine serum albumin Drugs 0.000 description 2
- 210000005013 brain tissue Anatomy 0.000 description 2
- 230000002490 cerebral effect Effects 0.000 description 2
- 238000010276 construction Methods 0.000 description 2
- 239000012470 diluted sample Substances 0.000 description 2
- 208000035475 disorder Diseases 0.000 description 2
- 238000001035 drying Methods 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 230000006870 function Effects 0.000 description 2
- 230000000302 ischemic effect Effects 0.000 description 2
- 239000006166 lysate Substances 0.000 description 2
- 238000013507 mapping Methods 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000010369 molecular cloning Methods 0.000 description 2
- 230000002107 myocardial effect Effects 0.000 description 2
- 230000017074 necrotic cell death Effects 0.000 description 2
- 230000008506 pathogenesis Effects 0.000 description 2
- 239000013612 plasmid Substances 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 239000013074 reference sample Substances 0.000 description 2
- 239000007790 solid phase Substances 0.000 description 2
- 239000012089 stop solution Substances 0.000 description 2
- 239000013598 vector Substances 0.000 description 2
- WBSMIPAMAXNXFS-UHFFFAOYSA-N 5-Nitro-2-(3-phenylpropylamino)benzoic acid Chemical compound OC(=O)C1=CC([N+]([O-])=O)=CC=C1NCCCC1=CC=CC=C1 WBSMIPAMAXNXFS-UHFFFAOYSA-N 0.000 description 1
- BSFODEXXVBBYOC-UHFFFAOYSA-N 8-[4-(dimethylamino)butan-2-ylamino]quinolin-6-ol Chemical compound C1=CN=C2C(NC(CCN(C)C)C)=CC(O)=CC2=C1 BSFODEXXVBBYOC-UHFFFAOYSA-N 0.000 description 1
- 102100027165 Alpha-2-macroglobulin receptor-associated protein Human genes 0.000 description 1
- 102100022977 Antithrombin-III Human genes 0.000 description 1
- 102100036451 Apolipoprotein C-I Human genes 0.000 description 1
- 102100030970 Apolipoprotein C-III Human genes 0.000 description 1
- 102000004152 Bone morphogenetic protein 1 Human genes 0.000 description 1
- 108090000654 Bone morphogenetic protein 1 Proteins 0.000 description 1
- 238000009010 Bradford assay Methods 0.000 description 1
- 208000014644 Brain disease Diseases 0.000 description 1
- 241000283707 Capra Species 0.000 description 1
- 102100026548 Caspase-8 Human genes 0.000 description 1
- 102100028003 Catenin alpha-1 Human genes 0.000 description 1
- 102100028906 Catenin delta-1 Human genes 0.000 description 1
- 206010008088 Cerebral artery embolism Diseases 0.000 description 1
- 206010008132 Cerebral thrombosis Diseases 0.000 description 1
- 102100032919 Chromobox protein homolog 1 Human genes 0.000 description 1
- 102100032918 Chromobox protein homolog 5 Human genes 0.000 description 1
- 102100026099 Claudin domain-containing protein 1 Human genes 0.000 description 1
- 102100038642 Cleavage and polyadenylation specificity factor subunit 2 Human genes 0.000 description 1
- 102100038250 Cyclin-G2 Human genes 0.000 description 1
- 102100026865 Cyclin-dependent kinase 5 activator 1 Human genes 0.000 description 1
- 238000001712 DNA sequencing Methods 0.000 description 1
- 102100027700 DNA-directed RNA polymerase I subunit RPA2 Human genes 0.000 description 1
- 102100032501 Death-inducer obliterator 1 Human genes 0.000 description 1
- 238000009007 Diagnostic Kit Methods 0.000 description 1
- 102100031695 DnaJ homolog subfamily C member 2 Human genes 0.000 description 1
- 102100030801 Elongation factor 1-alpha 1 Human genes 0.000 description 1
- 102100023688 Eotaxin Human genes 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- 241001198387 Escherichia coli BL21(DE3) Species 0.000 description 1
- 102100037733 Fatty acid-binding protein, brain Human genes 0.000 description 1
- 102100020760 Ferritin heavy chain Human genes 0.000 description 1
- 102100031351 Galectin-9 Human genes 0.000 description 1
- 102100028652 Gamma-enolase Human genes 0.000 description 1
- 102100030943 Glutathione S-transferase P Human genes 0.000 description 1
- 102100040352 Heat shock 70 kDa protein 1A Human genes 0.000 description 1
- 101000836956 Homo sapiens Alpha-2-macroglobulin receptor-associated protein Proteins 0.000 description 1
- 101000757319 Homo sapiens Antithrombin-III Proteins 0.000 description 1
- 101000928628 Homo sapiens Apolipoprotein C-I Proteins 0.000 description 1
- 101000793223 Homo sapiens Apolipoprotein C-III Proteins 0.000 description 1
- 101000983528 Homo sapiens Caspase-8 Proteins 0.000 description 1
- 101000859063 Homo sapiens Catenin alpha-1 Proteins 0.000 description 1
- 101000916264 Homo sapiens Catenin delta-1 Proteins 0.000 description 1
- 101000797584 Homo sapiens Chromobox protein homolog 1 Proteins 0.000 description 1
- 101000797581 Homo sapiens Chromobox protein homolog 5 Proteins 0.000 description 1
- 101000912657 Homo sapiens Claudin domain-containing protein 1 Proteins 0.000 description 1
- 101000957590 Homo sapiens Cleavage and polyadenylation specificity factor subunit 2 Proteins 0.000 description 1
- 101000884216 Homo sapiens Cyclin-G2 Proteins 0.000 description 1
- 101000650600 Homo sapiens DNA-directed RNA polymerase I subunit RPA2 Proteins 0.000 description 1
- 101000869896 Homo sapiens Death-inducer obliterator 1 Proteins 0.000 description 1
- 101000845887 Homo sapiens DnaJ homolog subfamily C member 2 Proteins 0.000 description 1
- 101000920078 Homo sapiens Elongation factor 1-alpha 1 Proteins 0.000 description 1
- 101000978392 Homo sapiens Eotaxin Proteins 0.000 description 1
- 101001027674 Homo sapiens Fatty acid-binding protein, brain Proteins 0.000 description 1
- 101001002987 Homo sapiens Ferritin heavy chain Proteins 0.000 description 1
- 101001130151 Homo sapiens Galectin-9 Proteins 0.000 description 1
- 101001058231 Homo sapiens Gamma-enolase Proteins 0.000 description 1
- 101001010139 Homo sapiens Glutathione S-transferase P Proteins 0.000 description 1
- 101001037759 Homo sapiens Heat shock 70 kDa protein 1A Proteins 0.000 description 1
- 101000990902 Homo sapiens Matrix metalloproteinase-9 Proteins 0.000 description 1
- 101000831266 Homo sapiens Metalloproteinase inhibitor 4 Proteins 0.000 description 1
- 101000873851 Homo sapiens N(G),N(G)-dimethylarginine dimethylaminohydrolase 1 Proteins 0.000 description 1
- 101001128158 Homo sapiens Nanos homolog 2 Proteins 0.000 description 1
- 101001128156 Homo sapiens Nanos homolog 3 Proteins 0.000 description 1
- 101000928278 Homo sapiens Natriuretic peptides B Proteins 0.000 description 1
- 101001124309 Homo sapiens Nitric oxide synthase, endothelial Proteins 0.000 description 1
- 101001124991 Homo sapiens Nitric oxide synthase, inducible Proteins 0.000 description 1
- 101000979629 Homo sapiens Nucleoside diphosphate kinase A Proteins 0.000 description 1
- 101001124867 Homo sapiens Peroxiredoxin-1 Proteins 0.000 description 1
- 101000728117 Homo sapiens Plasma membrane calcium-transporting ATPase 4 Proteins 0.000 description 1
- 101000612397 Homo sapiens Prenylcysteine oxidase 1 Proteins 0.000 description 1
- 101000611643 Homo sapiens Protein phosphatase 1 regulatory subunit 15A Proteins 0.000 description 1
- 101001092206 Homo sapiens Replication protein A 32 kDa subunit Proteins 0.000 description 1
- 101000963987 Homo sapiens SH3 domain-binding protein 5 Proteins 0.000 description 1
- 101000825071 Homo sapiens Sclerostin domain-containing protein 1 Proteins 0.000 description 1
- 101000800055 Homo sapiens Testican-1 Proteins 0.000 description 1
- 101000763314 Homo sapiens Thrombomodulin Proteins 0.000 description 1
- 101000622304 Homo sapiens Vascular cell adhesion protein 1 Proteins 0.000 description 1
- 101000955110 Homo sapiens WD repeat-containing protein 36 Proteins 0.000 description 1
- 108010001336 Horseradish Peroxidase Proteins 0.000 description 1
- 206010021143 Hypoxia Diseases 0.000 description 1
- 101710140350 Inactive dihydropteroate synthase 2 Proteins 0.000 description 1
- 201000001429 Intracranial Thrombosis Diseases 0.000 description 1
- 208000004552 Lacunar Stroke Diseases 0.000 description 1
- 206010051078 Lacunar infarction Diseases 0.000 description 1
- 108010016113 Matrix Metalloproteinase 1 Proteins 0.000 description 1
- 102100030412 Matrix metalloproteinase-9 Human genes 0.000 description 1
- 102100024289 Metalloproteinase inhibitor 4 Human genes 0.000 description 1
- 101100013460 Mus musculus Ftl2 gene Proteins 0.000 description 1
- 102100035854 N(G),N(G)-dimethylarginine dimethylaminohydrolase 1 Human genes 0.000 description 1
- 102100031893 Nanos homolog 3 Human genes 0.000 description 1
- 102100036836 Natriuretic peptides B Human genes 0.000 description 1
- 108010025020 Nerve Growth Factor Proteins 0.000 description 1
- 108010008858 Nitric Oxide Synthase Type I Proteins 0.000 description 1
- 102100022397 Nitric oxide synthase, brain Human genes 0.000 description 1
- 102100029438 Nitric oxide synthase, inducible Human genes 0.000 description 1
- 102100023252 Nucleoside diphosphate kinase A Human genes 0.000 description 1
- 102100037499 Parkinson disease protein 7 Human genes 0.000 description 1
- 208000031481 Pathologic Constriction Diseases 0.000 description 1
- 102100029139 Peroxiredoxin-1 Human genes 0.000 description 1
- 102100029743 Plasma membrane calcium-transporting ATPase 4 Human genes 0.000 description 1
- 108010064218 Poly (ADP-Ribose) Polymerase-1 Proteins 0.000 description 1
- 102100023712 Poly [ADP-ribose] polymerase 1 Human genes 0.000 description 1
- 229920001213 Polysorbate 20 Polymers 0.000 description 1
- 102100041004 Prenylcysteine oxidase 1 Human genes 0.000 description 1
- 108010032428 Protein Deglycase DJ-1 Proteins 0.000 description 1
- 102100029812 Protein S100-A12 Human genes 0.000 description 1
- 102100040714 Protein phosphatase 1 regulatory subunit 15A Human genes 0.000 description 1
- 102100040119 SH3 domain-binding protein 5 Human genes 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 102100022432 Sclerostin domain-containing protein 1 Human genes 0.000 description 1
- 208000006011 Stroke Diseases 0.000 description 1
- 102100033390 Testican-1 Human genes 0.000 description 1
- 102100026966 Thrombomodulin Human genes 0.000 description 1
- 102100023543 Vascular cell adhesion protein 1 Human genes 0.000 description 1
- 102100038944 WD repeat-containing protein 36 Human genes 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000001154 acute effect Effects 0.000 description 1
- 238000001042 affinity chromatography Methods 0.000 description 1
- 125000003275 alpha amino acid group Chemical group 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 230000000903 blocking effect Effects 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 230000008499 blood brain barrier function Effects 0.000 description 1
- 210000001218 blood-brain barrier Anatomy 0.000 description 1
- 210000004556 brain Anatomy 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 210000001175 cerebrospinal fluid Anatomy 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 238000011097 chromatography purification Methods 0.000 description 1
- 238000003776 cleavage reaction Methods 0.000 description 1
- 239000002299 complementary DNA Substances 0.000 description 1
- 230000002596 correlated effect Effects 0.000 description 1
- 230000000875 corresponding effect Effects 0.000 description 1
- 230000034994 death Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000003748 differential diagnosis Methods 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 230000004064 dysfunction Effects 0.000 description 1
- 238000001976 enzyme digestion Methods 0.000 description 1
- 102000052116 epidermal growth factor receptor activity proteins Human genes 0.000 description 1
- 108700015053 epidermal growth factor receptor activity proteins Proteins 0.000 description 1
- 239000013613 expression plasmid Substances 0.000 description 1
- 239000013604 expression vector Substances 0.000 description 1
- 108020001507 fusion proteins Proteins 0.000 description 1
- 102000037865 fusion proteins Human genes 0.000 description 1
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 1
- 239000010931 gold Substances 0.000 description 1
- 229910052737 gold Inorganic materials 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 229940098197 human immunoglobulin g Drugs 0.000 description 1
- 230000007954 hypoxia Effects 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 201000010849 intracranial embolism Diseases 0.000 description 1
- 208000028867 ischemia Diseases 0.000 description 1
- 125000001449 isopropyl group Chemical group [H]C([H])([H])C([H])(*)C([H])([H])[H] 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 208000011977 language disease Diseases 0.000 description 1
- 210000004880 lymph fluid Anatomy 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- HOVAGTYPODGVJG-UHFFFAOYSA-N methyl beta-galactoside Natural products COC1OC(CO)C(O)C(O)C1O HOVAGTYPODGVJG-UHFFFAOYSA-N 0.000 description 1
- 239000002808 molecular sieve Substances 0.000 description 1
- YOHYSYJDKVYCJI-UHFFFAOYSA-N n-[3-[[6-[3-(trifluoromethyl)anilino]pyrimidin-4-yl]amino]phenyl]cyclopropanecarboxamide Chemical compound FC(F)(F)C1=CC=CC(NC=2N=CN=C(NC=3C=C(NC(=O)C4CC4)C=CC=3)C=2)=C1 YOHYSYJDKVYCJI-UHFFFAOYSA-N 0.000 description 1
- 108010064131 neuronal Cdk5 activator (p25-p35) Proteins 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 239000013610 patient sample Substances 0.000 description 1
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 description 1
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 238000003259 recombinant expression Methods 0.000 description 1
- 238000005215 recombination Methods 0.000 description 1
- 230000006798 recombination Effects 0.000 description 1
- 230000000284 resting effect Effects 0.000 description 1
- 238000012502 risk assessment Methods 0.000 description 1
- 210000003296 saliva Anatomy 0.000 description 1
- 239000012898 sample dilution Substances 0.000 description 1
- 230000007017 scission Effects 0.000 description 1
- URGAHOPLAPQHLN-UHFFFAOYSA-N sodium aluminosilicate Chemical compound [Na+].[Al+3].[O-][Si]([O-])=O.[O-][Si]([O-])=O URGAHOPLAPQHLN-UHFFFAOYSA-N 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 1
- 230000036262 stenosis Effects 0.000 description 1
- 208000037804 stenosis Diseases 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 210000002700 urine Anatomy 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
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6893—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids related to diseases not provided for elsewhere
-
- 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
- G01N33/68—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
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/28—Neurological disorders
- G01N2800/2871—Cerebrovascular disorders, e.g. stroke, cerebral infarct, cerebral haemorrhage, transient ischemic event
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/52—Predicting or monitoring the response to treatment, e.g. for selection of therapy based on assay results in personalised medicine; Prognosis
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Immunology (AREA)
- Molecular Biology (AREA)
- Chemical & Material Sciences (AREA)
- Biomedical Technology (AREA)
- Urology & Nephrology (AREA)
- Hematology (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Cell Biology (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Peptides Or Proteins (AREA)
Abstract
本发明属于生物检测领域,具体涉及一种含有ICAM1蛋白片段的脑梗塞生物标记物及其应用。具体技术方案为:一种脑梗塞生物标记,包括与脑梗塞相关的抗原蛋白片段,至少包括:ICAM1蛋白片段。所述ICAM1蛋白片段的序列如SEQ ID NO.8。本发明提供的抗原或其组合可用于测试来自受试者的生物学样品中针对上述抗原组合的自身抗体的存在或存在水平,从而高效、简便地评估所述受试者患有脑梗塞的风险。
Description
技术领域
本发明属于生物检测领域,具体涉及一种含有ICAM1蛋白片段的脑梗塞生物标记物及其应用。
背景技术
脑梗塞,又称缺血性脑卒中(cerebral ischemic stroke),是指因脑部血液供应障碍,缺血、缺氧所导致的局部性脑组织的缺血性坏死或软化。脑梗塞的临床常见类型有脑血栓形成、腔隙性梗死和脑栓塞等,脑梗塞占全部脑卒中的80%。脑梗塞作为一种突发性脑疾病可以发生在任何年龄,坏死程度随发生部位及大小不同而有差别。常见于45~70岁中老年人,发病急,多无前驱症状,极易导致死亡。治疗后容易留下严重的后遗症,易复发,严重的出现肢体功能障碍,语言障碍和智力障碍。脑梗塞不仅给人类健康和生命造成极大威胁,而且给患者、家庭及社会带来极大的痛苦和沉重的负担。
脑梗塞的黄金治疗时间为发病后1~3小时以内。及时诊断和治疗对降低治疗后遗症,减少病人死亡率具有决定性意义。目前对脑梗塞的诊断方式主要是临床症状,结合CT、MRI等检查方法。然而,脑梗塞起病4~6小时内,只有部分病例可见边界不清的稍低密度灶,而大部分的病例在24小时后才能显示边界较清的低密度灶,已经远远超过了治疗的黄金窗口期。即:现有检查手段都需要发病后较长时间才能检测到病灶,容易延误病情,给病人带来严重的后遗症。因此,临床急需能够快速检查甚至提前预警脑梗塞风险的有效工具。
然而,探索能够快速诊断脑梗塞的生物标志物,特别是便捷的血清标志物,仍然是一项艰巨的任务。有研究人员利用脑梗塞病人血清中脑部应急反应产生的代谢产物或血清中相关抗原蛋白的浓度变化来研究它们与脑梗塞的相关性。这些标志物与脑梗塞都有一定的相关性。但是也存在明显的不足:多数标志物特异性不理想,在很多其它疾病中也广泛存在;部分标志物分子较大,不能及时、有效地通过血脑屏障。而且,由于脑梗塞发病的原因多种多样,具体的机理也不明确,单一的自身抗体指标无法有效覆盖各种病例。因此,现有标志物尚无法直接在临床上进行应用。
发明内容
本发明的目的是提供一种脑梗塞生物标记物及其应用。
为实现上述发明目的,本发明所采用的技术方案是:一种脑梗塞生物标记,包括与脑梗塞相关的抗原蛋白片段,至少包括:ICAM1蛋白片段和/或MMP1蛋白片段。
优选的,所述ICAM1蛋白片段的序列如SEQ ID NO.8;和/或,所述MMP1蛋白片段的序列如SEQ ID NO.2所示。
优选的,包括BDNF蛋白片段和ICAM1蛋白片段。
优选的,所述BDNF蛋白片段的序列如SEQ ID NO.5所示。
优选的,包括NF155蛋白片段,所述NF155蛋白片段的序列如SEQ ID NO.7所示。
优选的,包括NMDAR2A蛋白片段,所述NMDAR2A蛋白片段的序列如SEQ ID NO.9所示。
优选的,包括PRL蛋白片段和PDCD11蛋白片断,所述PRL蛋白片段的序列如SEQ IDNO.6所示,所述PDCD11蛋白片断的序列如SEQ ID NO.10所示。
优选的,包括NF155蛋白片段、NMDAR2A蛋白片段、NMDAR2B蛋白片段、PRL蛋白片段、PDCD11蛋白片段和LIMS4蛋白片段,所述NF155蛋白片段序列如SEQ ID NO.7所示,所述NMDAR2A蛋白片段序列如SEQ ID NO.9所示,所述NMDAR2B蛋白片段序列如SEQ ID NO.3所示,所述PRL蛋白片段序列如SEQ ID NO.6所示,所述PDCD11蛋白片断序列如SEQ ID NO.10所示,所述LIMS4蛋白片段序列如SEQ ID NO.1所示。
优选的,包括NF155蛋白片段、PRL蛋白片段、ICAM1蛋白片段、PDCD11蛋白片断和LIMS4蛋白片段和PUM3蛋白片段,所述NF155蛋白片段序列如SEQ ID NO.7所示,所述PRL蛋白片段序列如SEQ ID NO.6所示,所述ICAM1蛋白片段序列如SEQ ID NO.8所示,和/或,所述PDCD11蛋白片断序列如SEQ ID NO.10所示,所述LIMS4蛋白片段序列如SEQ ID NO.1所示,所述PUM3蛋白片段序列如SEQ ID NO.4所示。
相应的,利用所述脑梗塞生物标记制备的检测试纸、试剂、试剂盒。
本发明具有以下有益效果:本发明提供了一种用于检测脑梗塞相关自身抗体的蛋白质抗原或抗原组合物,其中涉及的蛋白质抗原或其片段可利用人工合成或基因重组的方法制得。例如,合成蛋白质抗原或其片段的编码DNA,以合成DNA为模板,设计引物,通过PCR、酶切、连接等分子克隆手段,将所述蛋白质抗原或其片段的基因片段克隆到表达质粒上,然后通过大肠杆菌、酵母或细胞进行表达,再经过层析纯化得到目标蛋白。同时,还可选择性地在蛋白抗原或其片段上增加GST、AVI、HIS、c-myc等标签。增加这些标签可以方便的对蛋白质抗原进行提纯或标记,但本质上并不会改变抗原与其自身抗体的结合特性。
本发明提供的抗原或其组合可用于测试来自受试者的生物学样品中针对上述抗原组合的自身抗体的存在或存在水平,从而判断所述受试者是否患有脑梗塞。可用于预测所述受试者是否具有罹患脑梗塞的风险;评估所述受试者所患有的脑梗塞的进展;或者判断所述受试者是否具有再发生脑梗塞的风险,还可用于辅助鉴别脑出血与脑梗塞,指导临床药物的使用。其中,所述生物学样品可以是血清、血浆、全血、唾液、口腔粘膜拭子、尿液、淋巴液、脑脊液等。根据具体情况,所述生物学样品可经提取、稀释、富集等手段进行预处理。使用方法多样、简便易操作。在将本发明提供的抗原组合用于上述测试时,通过使所述抗原组合中的蛋白质或其片段与可能存在的相应自身抗体发生结合或相互作用,来测试该自身抗体的存在或存在水平。
本发明提供的蛋白质抗原或其组合还可用于制备脑梗塞相关自身抗体检测试剂或脑梗塞诊断试剂。应当理解的,所述蛋白质抗原组合还可用于制备脑梗塞相关自身抗体检测试剂盒,所述试剂盒可以参照本发明实施例中使用所述蛋白质抗原组合进行脑梗塞相关自身抗体检测时的方法和试剂制备,也可根据需要进行相应调整。
综上,本发明提供了一种蛋白质抗原组合,其可用于脑梗塞的检测或诊断,尤其适用于脑梗塞发病前的风险评估和预测;并可根据需要进一步制备为相关试剂或试剂盒。
具体实施方式
下面对本发明实施例中的技术方案进行清楚、完整地描述,显然,所描述的实施例仅是本发明一部分实施例,而不是全部的实施例。若未特别指明,实施例中所用的技术手段为本领域技术人员所熟知的常规手段。所获得的数据均为进行至少3次重复后获得的平均值,且各重复获得的均为有效数据。
实施例一:抗原的重组载体构建、表达和纯化
1、抗原的选择。选择表1所示与脑梗塞高度相关的抗原蛋白,其数据库ID具体如表1所示。需要说明的是,发明人团队并非只对如下抗原蛋白进行了相关的实验选择,而是出于篇幅考虑只展示了部分具有代表性的抗原蛋白。另外,与脑梗塞相关的抗原蛋白非常多,并非选择任意相关蛋白均可有效鉴别脑梗塞;发明人团队有自己独有的筛选方法,出于商业秘密考虑,本文对具体筛选方法不予公开。
表1待测蛋白的数据库ID对应表
2、抗原重组载体的构建和表达。以人类cDNA文库(购自Invitrogen公司)或者全基因合成DNA为模板,分别设计引物,通过PCR、酶切、连接等分子克隆手段,将所述蛋白的基因片段克隆到pET28质粒上。同时,在蛋白N-端增加HIS、c-myc等标签,形成融合蛋白。将得到的重组表达载体通过DNA测序鉴定,确认包含正确的蛋白基因片段。需要说明的是:添加标签只是为了方便识别和提取蛋白,并未对蛋白作为抗原时的功能产生决定性影响,使用时,不添加标签或根据需要添加其它标签或采取其他标记/识别手段均可。
将上述包含蛋白基因片段的重组质粒转化至大肠杆菌BL21(DE3)感受态细胞中,挑取克隆接种至LB培养基中,37℃摇床培养。当菌体密度达到OD600约为0.8时,降温至16℃,在每个LB培养基中加入0.1mM异丙基硫代-β-D-半乳糖苷(IPTG),诱导表达过夜,获得菌体。
3、抗原的纯化。离心收集诱导表达的所述菌体,用PBS漂洗两遍。用裂解液(每g菌体加5~10mL裂解液)重悬并分散菌体,冰浴,超声破碎菌体(超声功率200W,破碎5s,休息5s)。破碎后13000rpm,10℃,离心20分钟,取上清,经Ni柱亲和层析和分子筛层析两步纯化后,采用SDS-PAGE电泳分析,确认蛋白的分子量、纯度,用Bradford法测定浓度后,保存在-80℃备用。即获得纯化后的待测蛋白。
实施例二:从待测蛋白中筛选获得候选抗原
1、本实施例中使用的溶液和试剂如下:
(1)包被缓冲液为PBS缓冲液,pH7.4。其制备方法为:准确称取3.58g Na2HPO4·12H2O、0.23g KH2PO4·2H2O、0.2g KCl、8.0g NaCl,溶解于水中,加水定容至1L。
(2)封闭液/样本稀释液/抗体稀释液:将10g BSA(牛血清白蛋白)溶解于包被缓冲液中,加水定容至1L。
(3)洗涤液:现配现用,使用前在包被缓冲液中加入0.5%Tween20(V/V),pH=7.4。
(4)TMB显色剂,购自KPL公司。
(5)终止液:1M盐酸。
2、固相包被待测蛋白。用包被缓冲液将实施例一获得的纯化后的各待测蛋白稀释至5μg/mL,加至96孔板,每孔50μL,4℃包被过夜。次日倒掉溶液,甩干,用洗涤液洗三次,每次每孔200μL。然后每孔加入200μL封闭液,室温孵育1h后,将封闭液倒掉,甩干,再用洗涤液洗三次,每次每孔200μL,并再次甩干;获得位于96孔板中的固相包被的抗原。
3、加入待测样本。将待测人血清用样本稀释液稀释100倍后,加入所述含待测蛋白的96孔板,每孔加稀释后的待测样本50μL。随后将96孔板置于微孔板振荡仪上,室温振荡孵育1h;甩干,用洗涤液洗三次,每次每孔200μL,并再次甩干。
4、加入酶标二抗。将1.0mg/mL辣根过氧化物酶标记的重组羊抗人免疫球蛋白G抗体(购自Jackson ImmunoResearch Inc.)用抗体稀释液稀释20000倍后,加入步骤3处理后的96孔板中,每孔加50μL。随后将96孔板置于微孔板振荡仪上,室温振荡孵育0.5h,将板甩干,用洗涤液洗三次,每次每孔200μL,并再次甩干。
5、显色反应及光密度值读数。在步骤4处理后的96孔板中,每孔加50μL TMB显色剂,振荡15s,室温下避光反应15min,再加入50μL终止液;然后用酶标仪读取450nm波长的吸收值,获得每个待测样本的检测信号(S)。
6、敏感性和特异性分析。分别取47例阳性样本(确诊为脑梗塞患者的血清)和48例阴性样本(健康受试者血清),按上述方法(450nm波长的吸收值)测定每个样本的检测信号(S)。以阴性样本为阴性参考样本,计算所有阴性参考样本检测信号(S)的平均值(M)及标准差(SD),以M+3SD为Cut Off值。将检测信号(S)≥Cut Off值的样本(S≥M+3SD)定为阳性;将检测信号(S)<Cut Off值的样本(S<M+3SD)定为阴性。
基于样本阳性和阴性结果计算特异性和敏感性。其中,特异性是指健康受试者样本被正确地判定为阴性的比例,即在阴性样本中被正确地判定为阴性的数量除以阴性样本总数。敏感性是指脑梗塞患者样本被正确地判定为阳性的比例,即在阳性样本中被判定为阳性的数量除以阳性样本总数。通过计算得出使用每个被测蛋白作为抗原进行样本检测时的敏感性和特异性。结果如表2所示。
表2各被测蛋白作为抗原的敏感性和特异性展示表
蛋白名 | 敏感性 | 特异性 | 蛋白名 | 敏感性 | 特异性 |
RPA2 | 11% | 83% | VWF | 2% | 98% |
TUBB4B | 11% | 91% | NGF | 6% | 96% |
ATP2B4 | 2% | 98% | VCAM1 | 2% | 98% |
BMP1 | 5% | 91% | FTH1 | 2% | 100% |
MMP-1 | 9% | 96% | FTL | 2% | 100% |
DHPS | 2% | 96% | DDAH1 | 2% | 96% |
SH3BP5 | 4% | 94% | ENO2 | 2% | 98% |
PDCD11 | 6% | 100% | APOC1 | 2% | 100% |
CBX1 | 6% | 90% | APOC3 | 2% | 98% |
APOH | 9% | 96% | SERPINC1 | 2% | 98% |
CTNNA1 | 4% | 94% | FABP7 | 2% | 98% |
LRPAP1 | 4% | 96% | PARK7 | 2% | 100% |
SPOCK1 | 2% | 100% | NME1 | 2% | 100% |
LIMS4 | 9% | 96% | CBX5 | 4% | 94% |
PUM3 | 9% | 96% | CCL11 | 4% | 92% |
LGALS9 | 4% | 94% | EGFR | 2% | 94% |
PPP1R15A | 11% | 88% | S100A12 | 2% | 92% |
P3H4 | 6% | 100% | TIMP4 | 2% | 100% |
SOSTDC1 | 4% | 88% | PRL | 11% | 96% |
CTNND1 | 2% | 96% | PCYOX1 | 6% | 92% |
CLDND1 | 4% | 92% | DIDO1 | 6% | 96% |
CCNG2 | 4% | 94% | CPSF2 | 2% | 100% |
DNAJC2 | 6% | 88% | THBD | 2% | 100% |
NMDA2B | 9% | 96% | NF155 | 13% | 100% |
NMDA2A | 9% | 98% | NOS3 | 2% | 100% |
HSPA1A | 11% | 92% | NOS2 | 2% | 100% |
PRDX1 | 2% | 98% | NOS1 | 2% | 96% |
GSTP1 | 2% | 92% | SELE | 2% | 100% |
SELL | 2% | 92% | sICAM | 2% | 100% |
MT3 | 4% | 90% | CASP8 | 6% | 94% |
FADD | 13% | 100% | PARP1 | 2% | 100% |
AIFM1 | 9% | 96% | CDK5R1 | 6% | 92% |
AVP | 4% | 96% | EEF1A1 | 6% | 94% |
S100B | 2% | 98% | FN1 | 2% | 100% |
BDNF | 11% | 96% | SAA | 4% | 96% |
MMP9 | 2% | 100% | ICAM1 | 10% | 100% |
NPPB | 4% | 94% | WDR36 | 2% | 98% |
7、按下述标准:敏感性>8%且特异性>90%或敏感性>5%且特异性>98%,从上述各被测蛋白中选择筛选出敏感性和特异性较好的蛋白作为候选抗原。筛选结果如表3所示。
表3从被测蛋白中筛选出的候选抗原展示表
候选抗原 | 敏感性 | 特异性 | 候选抗原 | 敏感性 | 特异性 |
TUBB4B | 11% | 91% | BDNF | 11% | 96% |
MMP1 | 9% | 96% | PRL | 11% | 96% |
PDCD11 | 6% | 100% | NF155 | 13% | 100% |
LIMS4 | 9% | 96% | ICAM1 | 10% | 100% |
PUM3 | 9% | 96% | NMDAR2A | 9% | 98% |
P3H4 | 6% | 100% | FADD | 13% | 100% |
NMDAR2B | 9% | 96% | AIFM1 | 9% | 96% |
APOH | 9% | 96% | \ | \ | \ |
实施例三:候选抗原的活性位点筛选
对实施例二获得的所述候选抗原的氨基酸序列及其结构进行分析,经大量前期试验后,从各所述候选抗原中选择不同的序列片段和抗原表位。所选择的序列片段如表4所示。
按照实施例一的方法,对表4中各蛋白片段进行表达载体构建、表达和纯化,得到构建后的蛋白片段,再按实施例二的方法检测各构建后的蛋白片段作为抗原时的敏感性和特异性。与实施例一相同的是,此处虽然也在各蛋白片段中添加了一些标签,但添加标签的目的主要在于便于提取和识别,并未对蛋白片段作为抗原的功能产生实质性影响。使用添加标签前后的蛋白均可实现发明目的,实际使用时,本领域技术人员可以根据需要选择是否添加标签、及添加标签的种类。因此,本发明全文只提供了添加标签前的蛋白列。测试结果如表4所示。
表4各候选抗原上截取的序列片段及敏感性和特异性对应表
根据表4的结果,结合不同抗原在体内的分布,进一步筛选出10种蛋白质片段,作为区分脑梗塞患者和健康受试者的候选抗原。筛选结果如表5所示。
表5各候选抗原的敏感性和特异性对照表
实施例四:候选抗原组合检测脑梗塞样本的效果展示
1、根据表5的筛选结果,从10种所述候选蛋白抗原片段中选择形成不同的抗原组合,结果如表6所示。需要说明的是,发明人并非只进行了表6的组合试验;发明人经过大量前期试验后才获得表6中所示的各抗原组合,因篇幅限制,只选取了部分效果较优的组合。
表6各抗原组合对照表
组合 | 组合中包含的候选蛋白抗原 |
组合1 | BDNF,ICAM1 |
组合2 | BDNF,ICAM1,MMP1 |
组合3 | BDNF,ICAM1,MMP1,NF155 |
组合4 | BDNF,ICAM1,MMP1,NF155,NMDA2A |
组合5 | BDNF,ICAM1,MMP1,NF155,PRL,PDCD11 |
组合6 | NF155,MMP1,NMDAR2A,NMDAR2B,PRL,PDCD11,LIMS4 |
组合7 | NF155,MMP1,PRL,ICAM1,PDCD11,LIMS4,PUM3 |
2、取脑梗塞患者血清样本2564例,取正常的健康受试者血清样本3576例。参照实施例二的检测方法,分别使用表6中各组抗原组合对上述各样本进行敏感性和特异性检测。
抗原组合中,单个抗原的阳性和阴性的定义方法参照实施例二。抗原组合的整体敏感性和特异性定义方法如下:
对于一个抗原组合,当某个血清样本采用该组合中的任意一个抗原得到的检测信号为阳性检测信号时,该血清样本为阳性样本;否则该血清样本为阴性样本。
基于上述阳性和阴性定义方法,检测获得一个抗原组合检测得到的阳性和阴性结果,进而计算该抗原组合在患者样本中的敏感性。
其中,敏感性是指在脑梗塞患者样本总共被正确地判定为阳性的比例,即在所有脑梗塞患者样本中被判定为阳性的数量除以所有脑梗塞患者样本总数;特异性是指健康受试者样本被正确地判定为阴性的比例,即在健康受试者样本中,健康受试者样本中被判定为阴性的数量除以健康受试者样本总数。
各组抗原组合检测获得的敏感性和特异性结果如表7所示。
表7各抗原组合检测脑梗塞样本的敏感性和特异性结果对照表
组合 | 敏感性 | 特异性 |
组合1 | 55% | 98% |
组合2 | 64% | 96% |
组合3 | 72% | 96% |
组合4 | 77% | 96% |
组合5 | 85% | 96% |
组合6 | 72% | 96% |
组合7 | 74% | 96% |
需要说明的是:表7结果是对大量样本检测后获得的实际结果,并未对结果进行任何的选择、修饰,一定程度上可直接反应应用到临床上的结果。在后期应用时,还可根据需要,从本发明提供的理想候选抗原或抗原组合物中选择一种或多种,制作成诊断试剂盒。
实施例五:候选抗原组合区分脑出血和脑梗塞的效果展示
脑出血与脑梗塞这两种疾病的临床症状极为相似,区别诊断比较困难。但是两者的发病本质是不同的,需要采取的临床治疗措施也有很大差异。临床上如果出现误诊,则很可能会给病人带来无法挽回的后果甚至生命危险。因此,对两者进行有效鉴别和区分具有重要的临床意义。
取脑出血患者血清样本94例,取正常的健康受试者血清样本96例。参照实施例四的检测方法,分别使用表6中各组抗原组合对上述各样本进行敏感性和特异性检测。各组抗原组合检测获得的敏感性和特异性结果如表8所示。本实施例中,敏感性的定义为:在脑出血患者样本总共被正确地判定为阳性的比例,即在所有脑出血患者样本中被判定为阳性的数量除以所有脑出血患者样本总数;特异性是指健康受试者样本被正确地判定为阴性的比例,即在健康受试者样本中,健康受试者样本中被判定为阴性的数量除以健康受试者样本总数。
表8各抗原组合检测脑出血样本的敏感性和特异性结果对照表
组合 | 敏感性 | 特异性 |
组合1 | 6.3% | 97% |
组合2 | 8.3% | 97% |
组合3 | 9.4% | 97% |
组合4 | 10.4% | 97% |
组合5 | 9.4% | 97% |
组合6 | 8.3% | 97% |
组合7 | 10.4% | 97% |
对照表7和表8的结果,我们可以看出:7组抗原组合对脑梗塞样本的检出率很高,其中组合的5敏感性最高,达到85%;而各组合对脑出血样本的检出率都较低,最高仅为10.4%。因此,各抗原组合均可区分脑梗塞与脑出血。
实施例六:候选抗原组合区分心肌梗死和脑梗塞的效果展示
心肌梗塞和脑梗塞的发病机制原理基本相同,都是由于动脉血管狭窄或阻塞导致供血不足而出现心肌细胞或脑细胞缺血性损伤。因此,临床上要区分这两种疾病也存在一定难度。本发明通过大量筛选,获得与脑部组织具有高度相关性,而又与心肌组织相关性较低的抗原或其片断,从而实现对两种疾病的区分检测。
取心肌梗死患者血清样本96例,取正常的健康受试者血清样本94例。参照实施例四的检测方法,分别使用表6中各组抗原组合对上述各样本进行敏感性和特异性检测。各组抗原组合检测获得的敏感性和特异性结果如表9所示。本实施例中,敏感性的定义为:在心肌梗死患者样本总共被正确地判定为阳性的比例,即在所有心肌梗死患者样本中被判定为阳性的数量除以所有心肌梗死患者样本总数;特异性是指健康受试者样本被正确地判定为阴性的比例,即在健康受试者样本中,健康受试者样本中被判定为阴性的数量除以健康受试者样本总数。
表9各抗原组合检测心肌梗死样本的敏感性和特异性结果对照表
对照表7和表9的结果,我们可以看出:各组合对心肌梗死样本的检出率都较低,最高仅为15.6%。因此,各抗原组合均可有效区分脑梗塞与心肌梗死。
以上所述的实施例仅是对本发明的优选方式进行描述,并非对本发明的范围进行限定,在不脱离本发明设计精神的前提下,本领域普通技术人员对本发明的技术方案做出的各种变形、变型、修改、替换,均应落入本发明权利要求书确定的保护范围内。
Claims (3)
1.一种脑梗塞生物标记,其特征在于:包括与脑梗塞相关的抗原蛋白片段,至少包括:ICAM1蛋白片段,所述ICAM1蛋白片段的序列如SEQ ID NO.8。
2.根据权利要求1所述脑梗塞生物标记,其特征在于:包括BDNF蛋白片段和ICAM1蛋白片段,所述BDNF蛋白片段的序列如SEQ ID NO.5所示。
3.利用权利要求1或2所述脑梗塞生物标记制备的检测试纸、试剂、试剂盒。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202210918772.8A CN116068191A (zh) | 2022-05-09 | 2022-05-09 | 一种含有icam1蛋白片段的脑梗塞生物标记物及其应用 |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202210918772.8A CN116068191A (zh) | 2022-05-09 | 2022-05-09 | 一种含有icam1蛋白片段的脑梗塞生物标记物及其应用 |
CN202210495578.3A CN114594273B (zh) | 2022-05-09 | 2022-05-09 | 一种脑梗塞生物标记物及其应用 |
Related Parent Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202210495578.3A Division CN114594273B (zh) | 2022-05-09 | 2022-05-09 | 一种脑梗塞生物标记物及其应用 |
Publications (1)
Publication Number | Publication Date |
---|---|
CN116068191A true CN116068191A (zh) | 2023-05-05 |
Family
ID=81821625
Family Applications (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202210495578.3A Active CN114594273B (zh) | 2022-05-09 | 2022-05-09 | 一种脑梗塞生物标记物及其应用 |
CN202210918772.8A Pending CN116068191A (zh) | 2022-05-09 | 2022-05-09 | 一种含有icam1蛋白片段的脑梗塞生物标记物及其应用 |
Family Applications Before (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202210495578.3A Active CN114594273B (zh) | 2022-05-09 | 2022-05-09 | 一种脑梗塞生物标记物及其应用 |
Country Status (1)
Country | Link |
---|---|
CN (2) | CN114594273B (zh) |
Family Cites Families (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20110287955A1 (en) * | 2008-08-15 | 2011-11-24 | National University Corporation Chiba University | Polypeptide marker for diagnosis of arteriosclerosis, method for detection of arteriosclerosis by using the maker or the like, and kit for diagnosis of arteriosclerosis |
WO2015009907A1 (en) * | 2013-07-17 | 2015-01-22 | The Johns Hopkins University | A multi-protein biomarker assay for brain injury detection and outcome |
JP6312302B2 (ja) * | 2014-01-06 | 2018-04-18 | 公益財団法人ヒューマンサイエンス振興財団 | 脳梗塞の診断マーカー |
CN107533048A (zh) * | 2015-02-05 | 2018-01-02 | 美国免疫阵列公司 | 用于诊断脑损伤或神经退行性变的方法和组合物 |
JP6725798B2 (ja) * | 2016-11-15 | 2020-07-22 | 藤倉化成株式会社 | 動脈硬化の検出方法 |
JP6942036B2 (ja) * | 2017-11-30 | 2021-09-29 | 藤倉化成株式会社 | 脳梗塞の発症リスクを高感度に検出する体液抗体バイオマーカー |
JP6499342B2 (ja) * | 2018-03-19 | 2019-04-10 | 隆樹 日和佐 | 脳梗塞の診断マーカー |
-
2022
- 2022-05-09 CN CN202210495578.3A patent/CN114594273B/zh active Active
- 2022-05-09 CN CN202210918772.8A patent/CN116068191A/zh active Pending
Also Published As
Publication number | Publication date |
---|---|
CN114594273B (zh) | 2024-02-27 |
CN114594273A (zh) | 2022-06-07 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Rodríguez-Suárez et al. | Urine as a source for clinical proteome analysis: from discovery to clinical application | |
KR101850827B1 (ko) | 인지기능장애질환의 바이오 마커 및 당해 바이오 마커를 사용하는 인지기능장애질환의 검출방법 | |
Wilson et al. | Antibody arrays in biomarker discovery | |
US20130203088A1 (en) | Methods and means for diagnosing spondylarthritis using autoantibody markers | |
JP2010510528A (ja) | 自己免疫疾患のバイオマーカー | |
KR20110104930A (ko) | 갈렉틴-3 면역검정 | |
EP2841945A1 (en) | Methods and compositions for diagnosis and prognosis of stroke or other cerebral injury | |
US9778250B2 (en) | Detecting inclusion body myositis | |
JP2013174442A (ja) | リウマチ障害と非リウマチ障害とを識別するためのトリガーアッセイ | |
JP2018205327A (ja) | 子癇前症を診断するための方法および組成物 | |
CN107110848B (zh) | 以脱氧羟腐胺缩赖氨酸合酶基因作为指标使用的动脉硬化及癌的检测方法 | |
US8697384B2 (en) | YKL-40 as a general marker for non-specific disease | |
CN114594273B (zh) | 一种脑梗塞生物标记物及其应用 | |
US20160161499A1 (en) | Sensitive diagnostic assay for inclusion body mysitis | |
KR102131860B1 (ko) | 아르기닌이 메틸화된 ggt1에 특이적으로 결합하는 대장암 진단용 바이오마커 조성물 | |
CN117538545B (zh) | 一种用于阿尔茨海默症检测的蛋白抗原组合及应用 | |
JP2009092561A (ja) | ジヒドロピリジン受容体抗体レベルに基づく胸腺腫合併重症筋無力症の診断方法 | |
CN114072677A (zh) | 癌症检测、预后和治疗监测的试剂和方法 | |
WO2024057956A1 (ja) | p53アイソフォーム変異体のがん診断用途 | |
CN111018973A (zh) | 一种抗人心肌肌钙蛋白i的重组抗体 | |
US8841084B2 (en) | Methods for the detection and monitoring of acute myocardial infarction | |
JP7523129B2 (ja) | ヘリコバクター・ピロリ菌株の同定方法、および同定用キット | |
JP6618107B2 (ja) | リンパ球性漏斗下垂体後葉炎検査試薬及びその用途 | |
CN109997043B (zh) | 护理点测定法 | |
KR101652894B1 (ko) | 대장암에 대한 신규 바이오마커 및 그의 용도 |
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 |