The application of isothermal amplification for detecting HBV-RNA in the monitoring of chronic hepatitis B patients

JIA Jia; CHEN Chu; Danzheng Jiancuo

doi:10.3969/j.issn.1007-8134.2025.04.005

Abstract：Objective To validate the methodological performance of the isothermal amplification technique for detecting hepatitis B virus(HBV)ribonucleic acid(RNA),and explore the application value of HBV RNA quantitative detection in the condition monitoring of chronic hepatitis B(CHB)patients.Methods Validation of the precision,lower limit of detection,linearity,and accuracy of RNA capture method for detecting HBV RNA.A total of 128 CHB patients diagnosed and hospitalized at the Shaanxi Provincial Hospital of Traditional Chinese Medicine from July to September 2022 were enrolled.All patients had"＜10 IU/mL"or"undetected"results in high-sensitivity HBV DNA quantitative tests.Simultaneous HBV RNA quantitative testing was conducted to analyze the relationship between HBV RNA levels,HBV DNA,HBeAg,and liver injury markers in the patients.Results The within-run precision for low and high values was 3.01%and 2.56%respectively,while the between-run precision was 3.74%and 2.60%respectively.The detection rate for samples close to the lower limit of detection,repeated 20 times,was 100%.The method demonstrated linearity within the range of HBV RNA concentration from 1.0×102 to 1.0×108 copies/mL(R2=0.996 7).The HBV RNA positivity rate in the group with HBV DNA quantitative results below the detection limit was 70.73%,while it was 43.68%in the group with undetected HBV DNA,showing a significant difference(P=0.004).There were also statistically significant differences in HBV RNA quantitative detection values between the 2 groups(P=0.000).The HBV RNA positivity rate in HBeAg-positive individuals was 89.47%,with an ALT abnormality rate of 57.89%.In HBeAg-negative individuals,the HBV RNA positivity rate was 45.87%,and the ALT abnormality rate was 49.54%.The difference in HBV RNA positivity rates between the 2 groups was statistically significant(P=0.000),while the difference in ALT abnormality rates was not statistically significant(P=0.502).The abnormal rate of ALT in HBV RNA-positive patients is 66.7%,compared to 37.1%in patients with negative HBV RNA detection,showing a significant difference(P=0.001).Conclusion The isothermal amplification method for detecting HBV RNA has reliable performance.It can still effectively detect viral RNA in treated CHB patients with HBV DNA below the detection limit,making it applicable for monitoring the condition of such patients.