Stability of Dried Blood Spots for HIV-1 Drug Resistance Analysis
| Autor: | Suzanne M. Crowe, Anna C. Hearps, Lisa M. Morris, Vicki Greengrass, Claire Ryan, Megan M. Plate |
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| Rok vydání: | 2010 |
| Předmět: |
Paper
Time Factors Genotype Human immunodeficiency virus (HIV) Drug resistance Biology medicine.disease_cause Sensitivity and Specificity behavioral disciplines and activities Cell Line Specimen Handling Plasma Virology Spin column-based nucleic acid purification Drug Resistance Viral medicine Humans Desiccation Dried blood High humidity Hematologic Tests Chromatography Spots Temperature Reproducibility of Results Humidity nervous system diseases Dried blood spot Blood surgical procedures operative Infectious Diseases nervous system HIV-1 therapeutics HIV drug resistance |
| Zdroj: | Current HIV Research. 8:134-140 |
| ISSN: | 1570-162X |
| DOI: | 10.2174/157016210790442740 |
| Popis: | The wide scale application of dried blood spots (DBS) as a collection tool for low-cost HIV drug resistance testing requires a greater understanding of the accuracy of DBS for genotype analysis and the stability of DBS under various environmental conditions. Analysis of a 50microl DBS via a single amplicon, nested PCR-based in-house assay (the Burnet genotyping assay) showed an average nucleotide concordance of 98.9% with plasma samples, although only 65% of nucleotide mixtures detected in plasma were also detected within DBS. The analysis of three DBS resulted in the detection of a greater number of nucleotide mixtures (72 and 109 mixtures detected within one and three DBS, respectively, n=10). Two DBS extraction protocols (silica particle; NucliSENS, bioMerieux and spin column extraction; High Pure, Roche) were assessed and found to be equivalent (79% and 84% recovery success respectively, n=19). FTA Elute paper (Whatman) was an inferior DBS collection medium compared to Whatman 903 paper. DBS appeared relatively tolerant to multiple freeze/thaw cycles, with 79% of DBS subjected to ten freeze/thaw cycles successfully amplified compared to 93% of DBS defrosted once (n=14). High temperature (37 degrees C) and high humidity (>90%) substantially impaired DBS recovery within two weeks of storage (38%, n=8), whilst storage at -20 degrees C or 4 degrees C adequately preserved DBS for this period (100% recovery, n=8). Therefore, whilst DBS are suitable for HIV drug resistance surveillance, the use of multiple DBS may be required to ensure accurate detection of minor HIV quasispecies and short-term storage of samples at either 4 degrees C or -20 degrees C is recommended. |
| Databáze: | OpenAIRE |
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