Sample-to-Answer Hepatitis B Virus DNA Detection from Whole Blood on a Centrifugal Microfluidic Platform with Double Rotation Axes.

Abstract

A point-of-care apparatus for hepatitis virus detection requires simple and easy-to-use processing steps and should have the same diagnostic capability as that in the central laboratory. However, no automated and efficient methods for hepatitis B virus (HBV) sample-to-answer detection include serum separation, and complete prestorage of reagents has been developed. We developed an automated sample-to-answer disc for rapid HBV detection from whole blood based on a double rotation axes centrifugal microfluidic platform. The disc with complete prestorage of reagents features fully automated and integrated serum separation from whole blood, magnetic bead-based DNA extraction, aliquoting of the nucleic acid, and real-time polymerase chain reaction. A laser diode for sequential release of prestored liquid reagents was used. Processing merely requires manual loading of the sample into the disc. We demonstrate successful sample-to-answer detection of HBV in a 500 μL whole blood sample with sample concentrations down to 102 copies/mL. The total time of the whole detection from sample-to-result is about 48 min. The disc provides a user-friendly molecular diagnostic system for rapid analysis of HBV without demanding a complicated laboratory instrument and major manual operation time. Overall, the results indicated that the developed disc could be used for HBV molecular diagnosis.