Simple detection of point mutations associated with HIV-1 drug resistance.
A novel assay is described for the detection of HIV-1 drug resistance that is simple, cheap and sensitive. HIV-1 drug resistance in B and non-B HIV-1 subtypes was investigated using Mutagenically-Separated PCR (MS--PCR) --- a competitive semi-nested PCR which uses mutagenic primers. The assay was as...
| Main Authors: | , , , , |
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| Format: | Journal article |
| Language: | English |
| Published: |
2001
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| _version_ | 1826287065660129280 |
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| author | Frater, A Chaput, C Beddows, S Weber, J McClure, M |
| author_facet | Frater, A Chaput, C Beddows, S Weber, J McClure, M |
| author_sort | Frater, A |
| collection | OXFORD |
| description | A novel assay is described for the detection of HIV-1 drug resistance that is simple, cheap and sensitive. HIV-1 drug resistance in B and non-B HIV-1 subtypes was investigated using Mutagenically-Separated PCR (MS--PCR) --- a competitive semi-nested PCR which uses mutagenic primers. The assay was assessed for sensitivity, specificity and its ability to detect mutant virus within a mixed mutant--wild-type population. Gene sequencing was carried out simultaneously for comparison. MS--PCR detected five copies of HIV-1 RNA from laboratory isolates and 50 copies from patient samples. We demonstrate 100% specificity of detection for wild type or mutant virus for clades A, B, C, D and E. For mixed populations of virus, MS--PCR can detect at least a 10% mix of wild type:mutant, or vice-versa. When applied to African patient samples MS--PCR detected 91.6% of the codons tested. Concordance with sequencing data was 88.8% for protease and 97.2% for RT. MS--PCR is sensitive and specific for the detection of mutations in HIV-1, and can be adapted easily to test for resistance at any codon of interest. |
| first_indexed | 2024-03-07T01:53:02Z |
| format | Journal article |
| id | oxford-uuid:9ac6b2c3-52cc-4c09-86ce-ec8a401df2a1 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-07T01:53:02Z |
| publishDate | 2001 |
| record_format | dspace |
| spelling | oxford-uuid:9ac6b2c3-52cc-4c09-86ce-ec8a401df2a12022-03-27T00:23:38ZSimple detection of point mutations associated with HIV-1 drug resistance.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:9ac6b2c3-52cc-4c09-86ce-ec8a401df2a1EnglishSymplectic Elements at Oxford2001Frater, AChaput, CBeddows, SWeber, JMcClure, MA novel assay is described for the detection of HIV-1 drug resistance that is simple, cheap and sensitive. HIV-1 drug resistance in B and non-B HIV-1 subtypes was investigated using Mutagenically-Separated PCR (MS--PCR) --- a competitive semi-nested PCR which uses mutagenic primers. The assay was assessed for sensitivity, specificity and its ability to detect mutant virus within a mixed mutant--wild-type population. Gene sequencing was carried out simultaneously for comparison. MS--PCR detected five copies of HIV-1 RNA from laboratory isolates and 50 copies from patient samples. We demonstrate 100% specificity of detection for wild type or mutant virus for clades A, B, C, D and E. For mixed populations of virus, MS--PCR can detect at least a 10% mix of wild type:mutant, or vice-versa. When applied to African patient samples MS--PCR detected 91.6% of the codons tested. Concordance with sequencing data was 88.8% for protease and 97.2% for RT. MS--PCR is sensitive and specific for the detection of mutations in HIV-1, and can be adapted easily to test for resistance at any codon of interest. |
| spellingShingle | Frater, A Chaput, C Beddows, S Weber, J McClure, M Simple detection of point mutations associated with HIV-1 drug resistance. |
| title | Simple detection of point mutations associated with HIV-1 drug resistance. |
| title_full | Simple detection of point mutations associated with HIV-1 drug resistance. |
| title_fullStr | Simple detection of point mutations associated with HIV-1 drug resistance. |
| title_full_unstemmed | Simple detection of point mutations associated with HIV-1 drug resistance. |
| title_short | Simple detection of point mutations associated with HIV-1 drug resistance. |
| title_sort | simple detection of point mutations associated with hiv 1 drug resistance |
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