Dot-blot hybridisation assay for detection of parvovirus B19 infections using synthetic oligonucleotides.
Pools of 10 synthetic oligonucleotides with sequences derived from the genome of parvovirus B19 and of 30 bases in length were made and labelled at the time of synthesis with digoxigenin (DIG) or dinitrophenyl (DNP) at the 5' end. They were used in a dot-blot hybridisation assay to detect parvo...
| Main Authors: | , , , , |
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| Format: | Journal article |
| Language: | English |
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1995
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| _version_ | 1797098891551703040 |
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| author | Cubie, H Molyneaux, P Shearman, M Gryzbowski, J Brown, T |
| author_facet | Cubie, H Molyneaux, P Shearman, M Gryzbowski, J Brown, T |
| author_sort | Cubie, H |
| collection | OXFORD |
| description | Pools of 10 synthetic oligonucleotides with sequences derived from the genome of parvovirus B19 and of 30 bases in length were made and labelled at the time of synthesis with digoxigenin (DIG) or dinitrophenyl (DNP) at the 5' end. They were used in a dot-blot hybridisation assay to detect parvovirus B19 DNA in sera submitted for routine virological diagnosis where parvovirus infection was suspected. Detection down to 10-100 fg DNA (equivalent to 10(3)-10(4) copies of parvovirus B19 genome) was obtained with both probe cocktails and colorimetric or chemiluminescent detection systems. Of 141 clinical samples examined from 126 patients presenting with rash and/or joint pains, 107 were clearly negative with both probes, 20 were clearly positive and the remaining 14 samples gave discrepant results. Of these 34 samples, 33 contained parvovirus B19 specific IgM. The parvovirus oligonucleotide probe cocktail produced and labelled with either DIG or DNP provided a useful diagnostic reagent for the detection of specific DNA in clinical specimens using a simple and sensitive dot-blot assay. |
| first_indexed | 2024-03-07T05:16:10Z |
| format | Journal article |
| id | oxford-uuid:dd43d978-bc5c-4c33-9278-f509feb0c134 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-07T05:16:10Z |
| publishDate | 1995 |
| record_format | dspace |
| spelling | oxford-uuid:dd43d978-bc5c-4c33-9278-f509feb0c1342022-03-27T09:23:52ZDot-blot hybridisation assay for detection of parvovirus B19 infections using synthetic oligonucleotides.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:dd43d978-bc5c-4c33-9278-f509feb0c134EnglishSymplectic Elements at Oxford1995Cubie, HMolyneaux, PShearman, MGryzbowski, JBrown, TPools of 10 synthetic oligonucleotides with sequences derived from the genome of parvovirus B19 and of 30 bases in length were made and labelled at the time of synthesis with digoxigenin (DIG) or dinitrophenyl (DNP) at the 5' end. They were used in a dot-blot hybridisation assay to detect parvovirus B19 DNA in sera submitted for routine virological diagnosis where parvovirus infection was suspected. Detection down to 10-100 fg DNA (equivalent to 10(3)-10(4) copies of parvovirus B19 genome) was obtained with both probe cocktails and colorimetric or chemiluminescent detection systems. Of 141 clinical samples examined from 126 patients presenting with rash and/or joint pains, 107 were clearly negative with both probes, 20 were clearly positive and the remaining 14 samples gave discrepant results. Of these 34 samples, 33 contained parvovirus B19 specific IgM. The parvovirus oligonucleotide probe cocktail produced and labelled with either DIG or DNP provided a useful diagnostic reagent for the detection of specific DNA in clinical specimens using a simple and sensitive dot-blot assay. |
| spellingShingle | Cubie, H Molyneaux, P Shearman, M Gryzbowski, J Brown, T Dot-blot hybridisation assay for detection of parvovirus B19 infections using synthetic oligonucleotides. |
| title | Dot-blot hybridisation assay for detection of parvovirus B19 infections using synthetic oligonucleotides. |
| title_full | Dot-blot hybridisation assay for detection of parvovirus B19 infections using synthetic oligonucleotides. |
| title_fullStr | Dot-blot hybridisation assay for detection of parvovirus B19 infections using synthetic oligonucleotides. |
| title_full_unstemmed | Dot-blot hybridisation assay for detection of parvovirus B19 infections using synthetic oligonucleotides. |
| title_short | Dot-blot hybridisation assay for detection of parvovirus B19 infections using synthetic oligonucleotides. |
| title_sort | dot blot hybridisation assay for detection of parvovirus b19 infections using synthetic oligonucleotides |
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