Establishment and application of ERA real-time fluorescence method for rapid detection of hepatitis B virus
This study is to establish a real-time fluorescence detection method for rapidly detecting the hepatitis B virus using enzyme recombinase amplification(ERA)technology.The primers and probes were designed according to the conserved sequence of the poly-merase coding region(P region)of the hepatitis B virus,and the reaction conditions,such as primers,temperature,and fluorescent probe concentration,were optimized to establish the optimal reaction system of the ERA real-time fluorescence assay,which was fur-ther validated in terms of sensitivity,specificity,anti-jamming capacity,and clinical samples.The results showed that the ERA real-time fluorescence method could detect the presence of the hepatitis B virus within 20 minutes at 42℃,with a limit of detection of 102 cop-ies/μL.The other four viruses(hepatitis A virus,hepatitis C virus,Epstein-Barr virus,and Cytomegalovirus)did not show fluorescence amplification curves,indicating reasonable specificity and anti-interference ability.When compared to the real-time fluorescence PCR method,the ERA real-time fluorescence method for 58 hepatitis B virus clinical samples exhibited a sensitivity of 97.37%,a specificity of 100%,a negative predictive value of 95.24%,a positive predictive value of 100%,and Kappa value of 0.96.This study successfully established a simple,rapid,sensitive,specific,and low-cost method for early detection of hepatitis B virus to meet the need for rapid detection.
enzymatic recombinase amplification(ERA)technologyrapid detectionhepatitis B virusthe ERA real-time fluorescence methodearly diagnosis
万方数据
维普
