Highly sensitive and specific detection of hepatitis B virus DNA and drug resistance mutations utilizing the PCR-based CRISPR-Cas13a system

ObjectivesUndetectable or low-level hepatitis B virus (HBV) DNA and drug resistance mutations in patients may increase the risk of HBV transmission cause active viral replication other clinical problems. Here, we established a highly sensitive practical method for detection using polymerase chain reaction (PCR) -based CRISPR-Cas13a system (referred to as PCR-CRISPR) evaluated its capability samples.MethodsSpecific CRISPR RNAs (crRNAs) are designed YMDD (tyrosine-methionine-aspartate-aspartate) variant identification. The was detected 312 serum samples diagnosis quantification PCR (qPCR) PCR-CRISPR. Additionally, 424 testing were by qPCR, direct sequencing, our assay.ResultsUsing PCR-CRISPR, one copy per test with HBV-1 crRNA 15 min after amplification. Consistent results qPCR observed 302 samples, while remaining 10 detectable PCR-CRISPR droplet digital but not qPCR. diagnosed all 412 drug-resistant kit well 12 undetectable sequencing.ConclusionsWe developed novel specific mutations. One could be detected. This has wide application prospects early infection, monitoring treatment guidance.