کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
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6135024 | 1593494 | 2010 | 6 صفحه PDF | دانلود رایگان |
![عکس صفحه اول مقاله: Novel application of Locked Nucleic Acid chemistry for a Taqman assay for measuring diverse human immunodeficiency virus type 1 subtypes Novel application of Locked Nucleic Acid chemistry for a Taqman assay for measuring diverse human immunodeficiency virus type 1 subtypes](/preview/png/6135024.png)
There remains a need for sensitive and cost-effective assays to monitor therapy in human immunodeficiency virus type-1 (HIV-1) infection. However, the genetic diversity of HIV poses difficulties for traditional real-time PCR assays that require long oligonucleotides probes. LNA⢠probes may be useful in overcoming these limits to traditional probe design. A new application of LNA⢠chemistry in a Taqman assay applicable to a wide range of HIV-1 subtypes is described. This assay, based on a 13-mer LNA⢠probe that matches the majority of HIV-1 sequences in the Los Alamos database, exhibited a wide dynamic range (101-107 copies of HIV DNA), high sensitivity (limit of detection of 1 copy of HIV DNA in 105 cells), and broad applicability to a range of HIV-1 subtypes (including A, B, C, D, F, H, B/C, and A/E CRFs). Using the LNA⢠probe assay, HIV-1 DNA was detected in all dried blood spots (DBS) from treatment naïve HIV-1 positive Ugandan children, and HIV DNA levels significantly correlated with viral RNA levels in plasma (r = 0.765, p < 0.0001). This approach to Taqman probe design should be explored further for use in diagnosis and monitoring of HIV in resource-limited settings, especially where several subtypes co-circulate.
Journal: Journal of Virological Methods - Volume 170, Issues 1â2, December 2010, Pages 115-120