کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
6140750 | 1594255 | 2014 | 8 صفحه PDF | دانلود رایگان |

- High Resolution melt (HRM) Analysis can detect GâtoâA hypermutated DNA sequences.
- An algorithm was established to enable quantitative hypermutation analyses by HRM.
- HyperHRM is a new method capable of accurately quantifying GâtoâA mutations in DNA.
- HRM was used to profile the deamination intensity of APOBEC3 proteins on HIV.
- HRM is a highâthroughput method to detect hypermutated HIV sequences in patients.
Enzymatic deamination of cytidines in DNA is an intrinsic component of antibody maturation and retroviral resistance, but can also be a source of HIV drug resistance and cancer-causing mutations. Here, we developed a high-throughput method based on high resolution melt (HRM) analysis called HyperHRM that can screen genomic DNA for rare hypermutated proviral sequences and accurately quantify the number of C-to-T or G-to-A mutations in each sequence. We demonstrate the effectiveness of the approach by profiling in parallel the intensity of the DNA mutator activity of all seven human APOBEC3 proteins on the near full-length sequence of HIV-1 and the Moloney murine leukemia virus. Additionally, HRM was successfully used to identify hypermutated proviral sequences in peripheral blood mononuclear cells from an HIV-1 patient. These results exemplify the effectiveness of HRM-based approaches for hypermutation quantification and for the detection of hypermutated DNA sequences potentially associated with disease or retroviral drug resistance.
Journal: Virology - Volume 448, 5 January 2014, Pages 168-175