CA2405769A1 - A method of distinguishing between living and dead cells using viability probes - Google Patents
A method of distinguishing between living and dead cells using viability probes Download PDFInfo
- Publication number
- CA2405769A1 CA2405769A1 CA002405769A CA2405769A CA2405769A1 CA 2405769 A1 CA2405769 A1 CA 2405769A1 CA 002405769 A CA002405769 A CA 002405769A CA 2405769 A CA2405769 A CA 2405769A CA 2405769 A1 CA2405769 A1 CA 2405769A1
- Authority
- CA
- Canada
- Prior art keywords
- nucleic acid
- cells
- sample
- living
- probe
- 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.)
- Abandoned
Links
- 239000000523 sample Substances 0.000 title claims abstract description 186
- 238000000034 method Methods 0.000 title claims abstract description 79
- 230000035899 viability Effects 0.000 title claims abstract description 78
- 150000007523 nucleic acids Chemical class 0.000 claims abstract description 148
- 108020004707 nucleic acids Proteins 0.000 claims abstract description 145
- 102000039446 nucleic acids Human genes 0.000 claims abstract description 145
- GHUXAYLZEGLXDA-UHFFFAOYSA-N 8-azido-5-ethyl-6-phenylphenanthridin-5-ium-3-amine;bromide Chemical compound [Br-].C12=CC(N=[N+]=[N-])=CC=C2C2=CC=C(N)C=C2[N+](CC)=C1C1=CC=CC=C1 GHUXAYLZEGLXDA-UHFFFAOYSA-N 0.000 claims description 49
- 238000001514 detection method Methods 0.000 claims description 46
- 230000003321 amplification Effects 0.000 claims description 43
- 238000003199 nucleic acid amplification method Methods 0.000 claims description 43
- 102000016911 Deoxyribonucleases Human genes 0.000 claims description 24
- 108010053770 Deoxyribonucleases Proteins 0.000 claims description 24
- 238000009739 binding Methods 0.000 claims description 20
- 230000027455 binding Effects 0.000 claims description 19
- 238000012986 modification Methods 0.000 claims description 11
- 230000004048 modification Effects 0.000 claims description 11
- 102000004190 Enzymes Human genes 0.000 claims description 8
- 108090000790 Enzymes Proteins 0.000 claims description 8
- 108091034117 Oligonucleotide Proteins 0.000 claims description 8
- ZMMJGEGLRURXTF-UHFFFAOYSA-N ethidium bromide Chemical group [Br-].C12=CC(N)=CC=C2C2=CC=C(N)C=C2[N+](CC)=C1C1=CC=CC=C1 ZMMJGEGLRURXTF-UHFFFAOYSA-N 0.000 claims description 7
- 108090000765 processed proteins & peptides Proteins 0.000 claims description 6
- 229960005542 ethidium bromide Drugs 0.000 claims description 5
- 230000002441 reversible effect Effects 0.000 claims description 5
- 238000002372 labelling Methods 0.000 claims description 4
- 108020005187 Oligonucleotide Probes Proteins 0.000 claims description 3
- 230000000415 inactivating effect Effects 0.000 claims description 3
- 239000002751 oligonucleotide probe Substances 0.000 claims description 3
- ZJVOHJVSTORGLI-UHFFFAOYSA-N 5-Methylangelicin Chemical compound O1C(=O)C=CC2=C1C(C=CO1)=C1C=C2C ZJVOHJVSTORGLI-UHFFFAOYSA-N 0.000 claims description 2
- 230000006037 cell lysis Effects 0.000 claims description 2
- 230000000593 degrading effect Effects 0.000 claims 2
- 210000004027 cell Anatomy 0.000 description 160
- 108020004414 DNA Proteins 0.000 description 64
- 241000894006 Bacteria Species 0.000 description 31
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N EtOH Substances CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 19
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 18
- 238000006243 chemical reaction Methods 0.000 description 17
- 239000000047 product Substances 0.000 description 13
- 239000007787 solid Substances 0.000 description 13
- 241000589875 Campylobacter jejuni Species 0.000 description 12
- 239000000975 dye Substances 0.000 description 11
- 238000003556 assay Methods 0.000 description 10
- 230000000694 effects Effects 0.000 description 10
- 239000003795 chemical substances by application Substances 0.000 description 9
- 238000002474 experimental method Methods 0.000 description 9
- 239000011324 bead Substances 0.000 description 8
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 8
- 238000012408 PCR amplification Methods 0.000 description 7
- 210000000170 cell membrane Anatomy 0.000 description 7
- 230000007613 environmental effect Effects 0.000 description 7
- 238000012163 sequencing technique Methods 0.000 description 7
- 238000011282 treatment Methods 0.000 description 7
- 238000000926 separation method Methods 0.000 description 6
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 5
- 239000011543 agarose gel Substances 0.000 description 5
- 239000000872 buffer Substances 0.000 description 5
- 238000000684 flow cytometry Methods 0.000 description 5
- 235000013305 food Nutrition 0.000 description 5
- 230000000937 inactivator Effects 0.000 description 5
- 244000005700 microbiome Species 0.000 description 5
- 230000035699 permeability Effects 0.000 description 5
- 239000000126 substance Substances 0.000 description 5
- 241000588724 Escherichia coli Species 0.000 description 4
- TWRXJAOTZQYOKJ-UHFFFAOYSA-L Magnesium chloride Chemical compound [Mg+2].[Cl-].[Cl-] TWRXJAOTZQYOKJ-UHFFFAOYSA-L 0.000 description 4
- 238000004458 analytical method Methods 0.000 description 4
- 230000001580 bacterial effect Effects 0.000 description 4
- 238000010438 heat treatment Methods 0.000 description 4
- 238000011534 incubation Methods 0.000 description 4
- 230000002401 inhibitory effect Effects 0.000 description 4
- 239000012528 membrane Substances 0.000 description 4
- 230000003287 optical effect Effects 0.000 description 4
- 230000002829 reductive effect Effects 0.000 description 4
- 108091008146 restriction endonucleases Proteins 0.000 description 4
- 241000589876 Campylobacter Species 0.000 description 3
- 238000002944 PCR assay Methods 0.000 description 3
- 238000011529 RT qPCR Methods 0.000 description 3
- 102000006943 Uracil-DNA Glycosidase Human genes 0.000 description 3
- 108010072685 Uracil-DNA Glycosidase Proteins 0.000 description 3
- 241000700605 Viruses Species 0.000 description 3
- 238000000246 agarose gel electrophoresis Methods 0.000 description 3
- 230000006907 apoptotic process Effects 0.000 description 3
- 230000008901 benefit Effects 0.000 description 3
- 230000015556 catabolic process Effects 0.000 description 3
- 230000030833 cell death Effects 0.000 description 3
- 238000006731 degradation reaction Methods 0.000 description 3
- 230000004069 differentiation Effects 0.000 description 3
- 210000003527 eukaryotic cell Anatomy 0.000 description 3
- 239000012634 fragment Substances 0.000 description 3
- 238000009396 hybridization Methods 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 102000004196 processed proteins & peptides Human genes 0.000 description 3
- 108090000623 proteins and genes Proteins 0.000 description 3
- 230000009467 reduction Effects 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 241000894007 species Species 0.000 description 3
- 239000006228 supernatant Substances 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- 108020004465 16S ribosomal RNA Proteins 0.000 description 2
- 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 description 2
- 102000053602 DNA Human genes 0.000 description 2
- 102000016928 DNA-directed DNA polymerase Human genes 0.000 description 2
- 108010014303 DNA-directed DNA polymerase Proteins 0.000 description 2
- 238000010222 PCR analysis Methods 0.000 description 2
- JLCPHMBAVCMARE-UHFFFAOYSA-N [3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-hydroxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methyl [5-(6-aminopurin-9-yl)-2-(hydroxymethyl)oxolan-3-yl] hydrogen phosphate Polymers Cc1cn(C2CC(OP(O)(=O)OCC3OC(CC3OP(O)(=O)OCC3OC(CC3O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c3nc(N)[nH]c4=O)C(COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3CO)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cc(C)c(=O)[nH]c3=O)n3cc(C)c(=O)[nH]c3=O)n3ccc(N)nc3=O)n3cc(C)c(=O)[nH]c3=O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)O2)c(=O)[nH]c1=O JLCPHMBAVCMARE-UHFFFAOYSA-N 0.000 description 2
- 230000004913 activation Effects 0.000 description 2
- 238000013459 approach Methods 0.000 description 2
- 239000012472 biological sample Substances 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 230000001413 cellular effect Effects 0.000 description 2
- 238000007796 conventional method Methods 0.000 description 2
- 230000001351 cycling effect Effects 0.000 description 2
- SUYVUBYJARFZHO-RRKCRQDMSA-N dATP Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@H]1C[C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)OP(O)(O)=O)O1 SUYVUBYJARFZHO-RRKCRQDMSA-N 0.000 description 2
- SUYVUBYJARFZHO-UHFFFAOYSA-N dATP Natural products C1=NC=2C(N)=NC=NC=2N1C1CC(O)C(COP(O)(=O)OP(O)(=O)OP(O)(O)=O)O1 SUYVUBYJARFZHO-UHFFFAOYSA-N 0.000 description 2
- RGWHQCVHVJXOKC-SHYZEUOFSA-J dCTP(4-) Chemical compound O=C1N=C(N)C=CN1[C@@H]1O[C@H](COP([O-])(=O)OP([O-])(=O)OP([O-])([O-])=O)[C@@H](O)C1 RGWHQCVHVJXOKC-SHYZEUOFSA-J 0.000 description 2
- HAAZLUGHYHWQIW-KVQBGUIXSA-N dGTP Chemical compound C1=NC=2C(=O)NC(N)=NC=2N1[C@H]1C[C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)OP(O)(O)=O)O1 HAAZLUGHYHWQIW-KVQBGUIXSA-N 0.000 description 2
- 238000013461 design Methods 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 239000007850 fluorescent dye Substances 0.000 description 2
- MURGITYSBWUQTI-UHFFFAOYSA-N fluorescin Chemical compound OC(=O)C1=CC=CC=C1C1C2=CC=C(O)C=C2OC2=CC(O)=CC=C21 MURGITYSBWUQTI-UHFFFAOYSA-N 0.000 description 2
- 230000007274 generation of a signal involved in cell-cell signaling Effects 0.000 description 2
- ZJYYHGLJYGJLLN-UHFFFAOYSA-N guanidinium thiocyanate Chemical compound SC#N.NC(N)=N ZJYYHGLJYGJLLN-UHFFFAOYSA-N 0.000 description 2
- 229910052739 hydrogen Inorganic materials 0.000 description 2
- 239000001257 hydrogen Substances 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 229910001629 magnesium chloride Inorganic materials 0.000 description 2
- 239000003550 marker Substances 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 238000010369 molecular cloning Methods 0.000 description 2
- 239000003068 molecular probe Substances 0.000 description 2
- 238000012544 monitoring process Methods 0.000 description 2
- 230000017074 necrotic cell death Effects 0.000 description 2
- 238000007826 nucleic acid assay Methods 0.000 description 2
- 238000009928 pasteurization Methods 0.000 description 2
- 230000002186 photoactivation Effects 0.000 description 2
- 210000001236 prokaryotic cell Anatomy 0.000 description 2
- 238000011002 quantification Methods 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 230000009870 specific binding Effects 0.000 description 2
- 238000010186 staining Methods 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- MXLMTQWGSQIYOW-UHFFFAOYSA-N 3-methyl-2-butanol Chemical compound CC(C)C(C)O MXLMTQWGSQIYOW-UHFFFAOYSA-N 0.000 description 1
- BZTDTCNHAFUJOG-UHFFFAOYSA-N 6-carboxyfluorescein Chemical compound C12=CC=C(O)C=C2OC2=CC(O)=CC=C2C11OC(=O)C2=CC=C(C(=O)O)C=C21 BZTDTCNHAFUJOG-UHFFFAOYSA-N 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- 229920000936 Agarose Polymers 0.000 description 1
- 239000012103 Alexa Fluor 488 Substances 0.000 description 1
- 108090000672 Annexin A5 Proteins 0.000 description 1
- 102000004121 Annexin A5 Human genes 0.000 description 1
- 241000203069 Archaea Species 0.000 description 1
- 244000063299 Bacillus subtilis Species 0.000 description 1
- 235000014469 Bacillus subtilis Nutrition 0.000 description 1
- 208000035143 Bacterial infection Diseases 0.000 description 1
- 241001112695 Clostridiales Species 0.000 description 1
- 108010062580 Concanavalin A Proteins 0.000 description 1
- 241000195493 Cryptophyta Species 0.000 description 1
- 108020003215 DNA Probes Proteins 0.000 description 1
- 238000007399 DNA isolation Methods 0.000 description 1
- 239000003298 DNA probe Substances 0.000 description 1
- AHCYMLUZIRLXAA-SHYZEUOFSA-N Deoxyuridine 5'-triphosphate Chemical compound O1[C@H](COP(O)(=O)OP(O)(=O)OP(O)(O)=O)[C@@H](O)C[C@@H]1N1C(=O)NC(=O)C=C1 AHCYMLUZIRLXAA-SHYZEUOFSA-N 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 108060004795 Methyltransferase Proteins 0.000 description 1
- 102000016397 Methyltransferase Human genes 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 101710163270 Nuclease Proteins 0.000 description 1
- 108091028043 Nucleic acid sequence Proteins 0.000 description 1
- 108010066717 Q beta Replicase Proteins 0.000 description 1
- 102000006382 Ribonucleases Human genes 0.000 description 1
- 108010083644 Ribonucleases Proteins 0.000 description 1
- BQCADISMDOOEFD-UHFFFAOYSA-N Silver Chemical compound [Ag] BQCADISMDOOEFD-UHFFFAOYSA-N 0.000 description 1
- 108020004682 Single-Stranded DNA Proteins 0.000 description 1
- ULHRKLSNHXXJLO-UHFFFAOYSA-L Yo-Pro-1 Chemical compound [I-].[I-].C1=CC=C2C(C=C3N(C4=CC=CC=C4O3)C)=CC=[N+](CCC[N+](C)(C)C)C2=C1 ULHRKLSNHXXJLO-UHFFFAOYSA-L 0.000 description 1
- 239000000061 acid fraction Substances 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 150000001298 alcohols Chemical class 0.000 description 1
- 230000000692 anti-sense effect Effects 0.000 description 1
- 238000003782 apoptosis assay Methods 0.000 description 1
- 208000022362 bacterial infectious disease Diseases 0.000 description 1
- 244000052616 bacterial pathogen Species 0.000 description 1
- 239000003139 biocide Substances 0.000 description 1
- 239000006161 blood agar Substances 0.000 description 1
- 210000001124 body fluid Anatomy 0.000 description 1
- 239000010839 body fluid Substances 0.000 description 1
- 238000009835 boiling Methods 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 238000009924 canning Methods 0.000 description 1
- 210000000234 capsid Anatomy 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 239000006285 cell suspension Substances 0.000 description 1
- 210000002421 cell wall Anatomy 0.000 description 1
- 239000001913 cellulose Substances 0.000 description 1
- 229920002678 cellulose Polymers 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 210000001175 cerebrospinal fluid Anatomy 0.000 description 1
- 238000007385 chemical modification Methods 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 125000004122 cyclic group Chemical group 0.000 description 1
- 238000004163 cytometry Methods 0.000 description 1
- 230000003111 delayed effect Effects 0.000 description 1
- 238000004925 denaturation Methods 0.000 description 1
- 230000036425 denaturation Effects 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 239000003599 detergent Substances 0.000 description 1
- 238000002405 diagnostic procedure Methods 0.000 description 1
- 230000009977 dual effect Effects 0.000 description 1
- 230000002900 effect on cell Effects 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 230000007515 enzymatic degradation Effects 0.000 description 1
- 230000009088 enzymatic function Effects 0.000 description 1
- 230000001747 exhibiting effect Effects 0.000 description 1
- 210000000887 face Anatomy 0.000 description 1
- 244000078673 foodborn pathogen Species 0.000 description 1
- ZHNUHDYFZUAESO-UHFFFAOYSA-N formamide Substances NC=O ZHNUHDYFZUAESO-UHFFFAOYSA-N 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 238000001502 gel electrophoresis Methods 0.000 description 1
- 229910052736 halogen Inorganic materials 0.000 description 1
- 150000002367 halogens Chemical class 0.000 description 1
- -1 heat Chemical compound 0.000 description 1
- 238000005286 illumination Methods 0.000 description 1
- 230000001900 immune effect Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000011065 in-situ storage Methods 0.000 description 1
- 230000002779 inactivation Effects 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 230000002458 infectious effect Effects 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 238000011901 isothermal amplification Methods 0.000 description 1
- 230000002147 killing effect Effects 0.000 description 1
- 238000007834 ligase chain reaction Methods 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 230000002934 lysing effect Effects 0.000 description 1
- 210000004962 mammalian cell Anatomy 0.000 description 1
- 230000000877 morphologic effect Effects 0.000 description 1
- 239000013642 negative control Substances 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 239000011236 particulate material Substances 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- INAAIJLSXJJHOZ-UHFFFAOYSA-N pibenzimol Chemical compound C1CN(C)CCN1C1=CC=C(N=C(N2)C=3C=C4NC(=NC4=CC=3)C=3C=CC(O)=CC=3)C2=C1 INAAIJLSXJJHOZ-UHFFFAOYSA-N 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 230000005522 programmed cell death Effects 0.000 description 1
- XJMOSONTPMZWPB-UHFFFAOYSA-M propidium iodide Chemical compound [I-].[I-].C12=CC(N)=CC=C2C2=CC=C(N)C=C2[N+](CCC[N+](C)(CC)CC)=C1C1=CC=CC=C1 XJMOSONTPMZWPB-UHFFFAOYSA-M 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 210000001938 protoplast Anatomy 0.000 description 1
- 230000005180 public health Effects 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 238000007859 qualitative PCR Methods 0.000 description 1
- 239000000376 reactant Substances 0.000 description 1
- 238000003757 reverse transcription PCR Methods 0.000 description 1
- 238000012552 review Methods 0.000 description 1
- 229940016590 sarkosyl Drugs 0.000 description 1
- 108700004121 sarkosyl Proteins 0.000 description 1
- 238000005204 segregation Methods 0.000 description 1
- 239000006152 selective media Substances 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 229910052709 silver Inorganic materials 0.000 description 1
- 239000004332 silver Substances 0.000 description 1
- KSAVQLQVUXSOCR-UHFFFAOYSA-M sodium lauroyl sarcosinate Chemical compound [Na+].CCCCCCCCCCCC(=O)N(C)CC([O-])=O KSAVQLQVUXSOCR-UHFFFAOYSA-M 0.000 description 1
- 239000002689 soil Substances 0.000 description 1
- 239000007790 solid phase Substances 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
- ABZLKHKQJHEPAX-UHFFFAOYSA-N tetramethylrhodamine Chemical compound C=12C=CC(N(C)C)=CC2=[O+]C2=CC(N(C)C)=CC=C2C=1C1=CC=CC=C1C([O-])=O ABZLKHKQJHEPAX-UHFFFAOYSA-N 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 238000011269 treatment regimen Methods 0.000 description 1
- GPRLSGONYQIRFK-MNYXATJNSA-N triton Chemical compound [3H+] GPRLSGONYQIRFK-MNYXATJNSA-N 0.000 description 1
- 241001515965 unidentified phage Species 0.000 description 1
- 210000002700 urine Anatomy 0.000 description 1
- 230000003612 virological effect Effects 0.000 description 1
- 230000000007 visual effect Effects 0.000 description 1
- 239000011534 wash buffer Substances 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6809—Methods for determination or identification of nucleic acids involving differential detection
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Analytical Chemistry (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Immunology (AREA)
- Molecular Biology (AREA)
- Biotechnology (AREA)
- Biophysics (AREA)
- Physics & Mathematics (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| GB0008787.4 | 2000-04-10 | ||
| GBGB0008787.4A GB0008787D0 (en) | 2000-04-10 | 2000-04-10 | A method of cell detection |
| PCT/GB2001/001619 WO2001077379A2 (en) | 2000-04-10 | 2001-04-10 | A method of cell detection |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA2405769A1 true CA2405769A1 (en) | 2001-10-18 |
Family
ID=9889587
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA002405769A Abandoned CA2405769A1 (en) | 2000-04-10 | 2001-04-10 | A method of distinguishing between living and dead cells using viability probes |
Country Status (8)
| Country | Link |
|---|---|
| US (1) | US7262009B2 (enExample) |
| EP (1) | EP1272664A2 (enExample) |
| JP (1) | JP4340734B2 (enExample) |
| AU (2) | AU4671201A (enExample) |
| CA (1) | CA2405769A1 (enExample) |
| GB (1) | GB0008787D0 (enExample) |
| NO (1) | NO20024879L (enExample) |
| WO (1) | WO2001077379A2 (enExample) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US8993259B2 (en) | 2012-05-02 | 2015-03-31 | Charles River Laboratories, Inc. | Method of viability staining with membrane permeable fluorescent dye and membrane impermeable fluorescence quencher |
| US9709500B2 (en) | 2012-05-02 | 2017-07-18 | Charles River Laboratories, Inc. | Optical method for detecting viable microorganisms in a cell sample |
| US10324036B2 (en) | 2012-05-02 | 2019-06-18 | Charles River Laboratories, Inc. | Porous planar cell capture system |
Families Citing this family (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DK1769081T3 (en) * | 2004-07-01 | 2016-03-14 | Litron Lab Ltd | PROCESS FOR COUNTING OF MAMMALS CELL micronuclei WITH A HEAD WEIGHT OF DIFFERENTIAL STAINING OF micronuclei AND chromatin OF DEAD AND DYING CELLS |
| US8026079B2 (en) | 2005-07-21 | 2011-09-27 | Morinaga Milk Industry Co., Ltd. | Method for detection of microorganism and kit for detection of microorganism |
| CA2615089C (en) | 2006-02-17 | 2015-11-17 | Morinaga Milk Industry Co., Ltd. | Method for detection of microorganism and kit for detection of microorganism |
| EP2004843B1 (en) * | 2006-02-28 | 2015-07-22 | Montana State University | Use of phenanthridium derivatives for distinguishing between intact and membrane-compromised cells using molecular nucleic acid-based techniques |
| JP4113556B2 (ja) * | 2006-12-04 | 2008-07-09 | 国立大学法人九州大学 | 核酸分解剤および核酸の分解方法 |
| WO2009022558A1 (ja) | 2007-08-16 | 2009-02-19 | Kyushu University | 微生物検出法及び微生物検出キット |
| KR101383389B1 (ko) | 2009-07-24 | 2014-04-08 | 모리나가 뉴교 가부시키가이샤 | 미생물 검출법 및 미생물 검출 키트 |
| WO2011043737A1 (en) * | 2009-10-05 | 2011-04-14 | Nanyang Technological University | Viability analysis of protozoa using polymerase chain reaction (pcr) |
| FR2974117B1 (fr) * | 2011-04-18 | 2018-05-11 | Societe Des Eaux De Marseille | Procede de quantification de bacteries vivantes dans un milieu liquide |
| DE102020103971B4 (de) * | 2020-02-14 | 2022-02-24 | Testo bioAnalytics GmbH | Verfahren zum Nachweis von lebenden Mikroorganismen und ein fluidisches Kanalsystem |
| KR20230079375A (ko) | 2020-09-02 | 2023-06-07 | 프로메가 코포레이션 | 핵산 변형 시약 및 이의 용도 |
Family Cites Families (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0517154A1 (en) * | 1991-06-06 | 1992-12-09 | F. Hoffmann-La Roche Ag | Method for detecting giardia |
| US5314805A (en) * | 1991-10-28 | 1994-05-24 | Molecular Probes, Inc. | Dual-fluorescence cell viability assay using ethidium homodimer and calcein AM |
| US5525471A (en) * | 1994-10-12 | 1996-06-11 | The United States Of America As Represented By The Secretary Of The Department Of Health And Human Services | Enzymatic degrading subtraction hybridization |
| US20020119438A1 (en) | 1995-12-29 | 2002-08-29 | Fuminori Kato | Method for determining viable cell count |
| US5780278A (en) * | 1996-05-20 | 1998-07-14 | Vanderbilt University | IceA gene and related methods |
| GB9709728D0 (en) | 1997-05-13 | 1997-07-02 | Dynal As | Single step method |
-
2000
- 2000-04-10 GB GBGB0008787.4A patent/GB0008787D0/en not_active Ceased
-
2001
- 2001-04-10 US US10/257,493 patent/US7262009B2/en not_active Expired - Fee Related
- 2001-04-10 EP EP01919651A patent/EP1272664A2/en not_active Withdrawn
- 2001-04-10 WO PCT/GB2001/001619 patent/WO2001077379A2/en not_active Ceased
- 2001-04-10 JP JP2001575233A patent/JP4340734B2/ja not_active Expired - Lifetime
- 2001-04-10 AU AU4671201A patent/AU4671201A/xx active Pending
- 2001-04-10 CA CA002405769A patent/CA2405769A1/en not_active Abandoned
- 2001-04-10 AU AU2001246712A patent/AU2001246712C1/en not_active Ceased
-
2002
- 2002-10-09 NO NO20024879A patent/NO20024879L/no unknown
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US8993259B2 (en) | 2012-05-02 | 2015-03-31 | Charles River Laboratories, Inc. | Method of viability staining with membrane permeable fluorescent dye and membrane impermeable fluorescence quencher |
| US8993260B2 (en) | 2012-05-02 | 2015-03-31 | Charles River Laboratories, Inc. | Fluorescence-based viability staining method using a membrane permeable flourescent dye and membrane impermeable fluorescence quencher |
| US9709500B2 (en) | 2012-05-02 | 2017-07-18 | Charles River Laboratories, Inc. | Optical method for detecting viable microorganisms in a cell sample |
| US10324036B2 (en) | 2012-05-02 | 2019-06-18 | Charles River Laboratories, Inc. | Porous planar cell capture system |
| US10976258B2 (en) | 2012-05-02 | 2021-04-13 | Charles River Laboratories, Inc. | Porous planar cell capture system and method of use |
Also Published As
| Publication number | Publication date |
|---|---|
| NO20024879D0 (no) | 2002-10-09 |
| AU2001246712B2 (en) | 2006-10-19 |
| JP2003530118A (ja) | 2003-10-14 |
| NO20024879L (no) | 2002-12-10 |
| EP1272664A2 (en) | 2003-01-08 |
| US20030203374A1 (en) | 2003-10-30 |
| WO2001077379A2 (en) | 2001-10-18 |
| GB0008787D0 (en) | 2000-05-31 |
| JP4340734B2 (ja) | 2009-10-07 |
| AU4671201A (en) | 2001-10-23 |
| US7262009B2 (en) | 2007-08-28 |
| AU2001246712C1 (en) | 2007-08-02 |
| WO2001077379A3 (en) | 2002-06-20 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US11746389B2 (en) | Method and kit of detecting the absence of micro-organisms | |
| Wang et al. | Discrimination of viable Vibrio vulnificus cells from dead cells in real-time PCR | |
| US6699670B2 (en) | Quantitative assay for the simultaneous detection and speciation of bacterial infections | |
| EP3775254B1 (en) | Microorganism separation and detection | |
| AU2001246712B2 (en) | A method of cell detection | |
| AU2001246712A1 (en) | A method of cell detection | |
| JP2016538843A (ja) | 微生物を複雑な試料から単離する方法 | |
| CA2976934A1 (en) | Ion torrent genomic sequencing | |
| Boujelben et al. | A broad-range PCR technique for the diagnosis of infective endocarditis | |
| Hill et al. | The polymerase chain reaction in molecular and micro-biology | |
| US20260117313A1 (en) | A crispr rna for detection of burkholderia pseudomallei-associated genetic material in a biological sample and a method using the same | |
| Cerna-Cortes et al. | Isolation of Mycobacterium mucogenicum from street-vended chili sauces: a potential source of human infection | |
| Kondoh et al. | Comparison of arbitrarily primed‐polymerase chain reaction and pulse‐field gel electrophoresis for characterizing methicillin‐resistant Staphylococcus aureus | |
| JP2012187067A (ja) | ラクトバチルス属細菌の定量方法 | |
| US9738939B2 (en) | Method for differentiating between living and dead cells | |
| Gorde et al. | Soil DNA extraction techniques for soil health and soil security | |
| JPH06253847A (ja) | 炭疽菌検出のためのオリゴヌクレオチドおよびそれを用いた検出方法 | |
| EP4634405A1 (en) | Method for determining the number and/or concentration of germinable cells in a sample of biological origin | |
| JPH06261759A (ja) | 炭疽菌検出のためのオリゴヌクレオチドおよびそれを用いた検出方法 |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| EEER | Examination request | ||
| FZDE | Discontinued |
Effective date: 20121126 |