Rapid and facile detection of HBV with CRISPR/Cas13a

Abstract

Hepatitis B remains a major global public health challenge, with particularly high prevalence in resource-poor Western Pacific and African regions. The lack of specialized pathogen detection equipment, such as qPCR, results in low hepatitis B virus (HBV) detection rates in resource-poor areas, and hinders efforts to combat the high prevalence of chronic hepatitis B. Therefore, the development of a rapid and accurate point-of-care test is essential for the prevention and control of hepatitis B in medically underdeveloped rural areas. In recent years, the application of clustered regularly interspaced short palindromic repeat (CRISPR)/Cas systems for nucleic acid detection has brought a new solution for the point-of-care detection of pathogens. We have developed a rapid and accurate point-of-care assay based on CRISPR-Cas13a for HBV. A limit of detection as low as 1 copy per μL was achieved by the Cas13a assay within 50 min. The lateral flow test strip technology can achieve visible results with the naked eye, whereas the fluorescence readout can achieve real-time high-sensitivity detection. In the evaluation of 74 clinical samples, the sensitivity rates of the fluorescence readout and lateral flow test strip method were 93.8% and 90.6%, respectively. The CRISPR/Cas13a-based HBV detection platform not only relies on minimal equipment to provide rapid, accurate and handy test results at low cost but also provides significant practical value for potential point-of-care HBV detection in areas where medical equipment is limited and beyond.