The Establishment Of A Magnetic Beads Based Nucleic Acid Extraction Method, And Its Clinical Application For Quantitative Detection Of Hepatitis Virus

To develop a rapid, convenient and high throughput automatic nucleic acid extraction method based on magnetic beads, for the detection of HBV and HCV viral load in human blood. The method and its related diagnostic kit established are ideal for clinical use. Using hepatitis B virus DNA and hepatitis C virus RNA extraction as examples, to establish and evaluate this extraction method.1) The key components of extraction reagents were selected and optimized, including the type and quantity of magnetic beads, the GTC concentration of lysis buffer, pH, the combining and washing conditions of magnetic beads, etc.2) The clinical performance of this established kit was evaluated for its linear range, precision, sensitivity, specificity, interference and resistance to contamination.3) The comparison study was performed using commercially available kits (boiling method and Roche COBAS TaqMan Test) with this established kit for their linear regression. In addition, the internal standard and the external standard were used in the kit to quality control and monitor the whole process of the testing and quantitative detection of samples respectively. The results showed that the performance of this established kit has meet the requirement of quantitative detection of hepatitis virus in clinical diagnosis in terms of the linear range, precision, sensitivity, specificity, interference and resistance to contamination of the established kit. The magnetic beads based method established in this study provides a better way of nucleic acid extraction, with increased the sensitivity, specificity and linear range of the testing kit. The beads method is also more resistant to sample interference.