title: Extraction of HCV-RNA from Plasma Samples: Development towards Semiautomation.
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 × (10(6), 10(5), 10(4), and 10(3)) 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 10(2) 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.
authors: PCR Laboratory, Punjab Institute of Nuclear Medicine (PINUM), Faisalabad 2019, Pakistan.
authors: Imran
authors: I
authors: Amin
authors: PCR Laboratory, Punjab Institute of Nuclear Medicine (PINUM), Faisalabad 2019, Pakistan ; Department of Clinical Pathology, Punjab Institute of Nuclear Medicine (PINUM), Faisalabad 2019, Pakistan.
authors: Tania
authors: T
authors: Jabbar
authors: PCR Laboratory, Punjab Institute of Nuclear Medicine (PINUM), Faisalabad 2019, Pakistan.
authors: Fawad
authors: F
authors: Niazi
authors: Nuclear Medicine Department, Punjab Institute of Nuclear Medicine (PINUM), Faisalabad 2019, Pakistan.
authors: Muhammad Saeed
authors: MS
authors: Akhtar