Automated Centrifugal Microfluidic Chip Integrating Pretreatment and Molecular Diagnosis for Hepatitis B Virus Genotyping from Whole Blood.

Point-of-care (POC) testing for nucleic acid that combines pretreatment and molecular diagnosis is crucial in analyzing complex samples such as those encountered in clinical diagnosis. Herein, we developed a centrifugal microfluidic platform, which can achieve a series of functions including separating serum and adsorbing, washing, eluting, and detecting DNA. We combined multiple signal enhancement systems including recombinase polymerase amplification (RPA), T7 transcription technology, and clustered regularly interspaced short palindromic repeat (CRISPR) technology to yield an ultrabright signal, which can avoid false-negative results. As an application, hepatitis B virus (HBV), a virus that causes global public health problems, was successfully detected and genotyped from whole blood on the automated centrifugal microfluidic platform. Compared to the traditional diagnosis process, the POC platform largely decreased the consumption of time from 3 to 1 h and the consumption of professional labor from three persons to only one. The automated centrifugal microfluidic platform integrated pretreatment and molecular diagnosis will play an essential role in clinical detection.