Extraction of HCV-RNA from Plasma Samples: Development towards Semiautomation
A semiautomated extraction protocol of HCV-RNA using Favorgen RNA extraction kit has been developed. The kit provided protocol was modified by replacing manual spin steps with vacuum filtration. The assay performance was evaluated by real-time qPCR based on Taqman technology. Assay linearity was con...
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| Format: | Article |
| Language: | English |
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Wiley
2015-01-01
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| Series: | International Journal of Analytical Chemistry |
| Online Access: | http://dx.doi.org/10.1155/2015/367801 |
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| author | Imran Amin Tania Jabbar Fawad Niazi Muhammad Saeed Akhtar |
| author_facet | Imran Amin Tania Jabbar Fawad Niazi Muhammad Saeed Akhtar |
| author_sort | Imran Amin |
| collection | DOAJ |
| description | A semiautomated extraction protocol of HCV-RNA using Favorgen RNA extraction kit has been developed. The kit provided protocol was modified by replacing manual spin steps with vacuum filtration. The assay performance was evaluated by real-time qPCR based on Taqman technology. Assay linearity was confirmed with the serial dilutions of RTA (Turkey) containing 1 × (106, 105, 104, and 103) IU mL−1. Comparison of test results obtained by two extraction methods showed a good correlation (r=0.95, n=30) with detection limit of 102 IU mL−1. The semiautomated vacuum filtration based protocol demonstrated high throughput: 35 minutes for the extraction of a batch of 30 samples (150 µL each) with reduced labor, time, waste, and cost. Performance characteristics of semiautomated system make it suitable for use in diagnostic purpose and viral load determinations. |
| format | Article |
| id | doaj-art-96ecffe1ad874f2a9d7119ff7548bb76 |
| institution | OA Journals |
| issn | 1687-8760 1687-8779 |
| language | English |
| publishDate | 2015-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | International Journal of Analytical Chemistry |
| spelling | doaj-art-96ecffe1ad874f2a9d7119ff7548bb762025-08-20T02:09:49ZengWileyInternational Journal of Analytical Chemistry1687-87601687-87792015-01-01201510.1155/2015/367801367801Extraction of HCV-RNA from Plasma Samples: Development towards SemiautomationImran Amin0Tania Jabbar1Fawad Niazi2Muhammad Saeed Akhtar3PCR Laboratory, Punjab Institute of Nuclear Medicine (PINUM), Faisalabad 2019, PakistanPCR Laboratory, Punjab Institute of Nuclear Medicine (PINUM), Faisalabad 2019, PakistanPCR Laboratory, Punjab Institute of Nuclear Medicine (PINUM), Faisalabad 2019, PakistanNuclear Medicine Department, Punjab Institute of Nuclear Medicine (PINUM), Faisalabad 2019, PakistanA semiautomated extraction protocol of HCV-RNA using Favorgen RNA extraction kit has been developed. The kit provided protocol was modified by replacing manual spin steps with vacuum filtration. The assay performance was evaluated by real-time qPCR based on Taqman technology. Assay linearity was confirmed with the serial dilutions of RTA (Turkey) containing 1 × (106, 105, 104, and 103) IU mL−1. Comparison of test results obtained by two extraction methods showed a good correlation (r=0.95, n=30) with detection limit of 102 IU mL−1. The semiautomated vacuum filtration based protocol demonstrated high throughput: 35 minutes for the extraction of a batch of 30 samples (150 µL each) with reduced labor, time, waste, and cost. Performance characteristics of semiautomated system make it suitable for use in diagnostic purpose and viral load determinations.http://dx.doi.org/10.1155/2015/367801 |
| spellingShingle | Imran Amin Tania Jabbar Fawad Niazi Muhammad Saeed Akhtar Extraction of HCV-RNA from Plasma Samples: Development towards Semiautomation International Journal of Analytical Chemistry |
| title | Extraction of HCV-RNA from Plasma Samples: Development towards Semiautomation |
| title_full | Extraction of HCV-RNA from Plasma Samples: Development towards Semiautomation |
| title_fullStr | Extraction of HCV-RNA from Plasma Samples: Development towards Semiautomation |
| title_full_unstemmed | Extraction of HCV-RNA from Plasma Samples: Development towards Semiautomation |
| title_short | Extraction of HCV-RNA from Plasma Samples: Development towards Semiautomation |
| title_sort | extraction of hcv rna from plasma samples development towards semiautomation |
| url | http://dx.doi.org/10.1155/2015/367801 |
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