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International Journal of Analytical Chemistry
Volume 2015, Article ID 367801, 4 pages
http://dx.doi.org/10.1155/2015/367801
Research Article

Extraction of HCV-RNA from Plasma Samples: Development towards Semiautomation

1PCR Laboratory, Punjab Institute of Nuclear Medicine (PINUM), Faisalabad 2019, Pakistan
2Department of Clinical Pathology, Punjab Institute of Nuclear Medicine (PINUM), Faisalabad 2019, Pakistan
3Nuclear Medicine Department, Punjab Institute of Nuclear Medicine (PINUM), Faisalabad 2019, Pakistan

Received 30 September 2014; Revised 4 February 2015; Accepted 4 February 2015

Academic Editor: Mohammad R. Pourjavid

Copyright © 2015 Imran Amin et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

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 (, ) 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.