传染病信息

目的 验证恒温扩增技术检测乙型肝炎病毒(hepatitis B virus, HBV)核糖核酸(ribonucleic acid, RNA)的方法学性能，并探讨HBV RNA定量检测在慢性乙型肝炎(chronic hepatitis B, CHB)患者病情监测中的应用价值。方法 对RNA捕获法检测HBV RNA进行精密度、最低检出限、线性和准确性的性能验证。纳入2022年7—9月在陕西省中医医院确诊并住院治疗的CHB患者共计128例，高敏HBV脱氧核糖核酸(deoxyribonucleic acid, DNA)定量检测结果均为“<10IU/mL”或者“未检出”，同时进行HBV RNA定量检测，分析患者HBV RNA水平与HBV DNA、HBeAg以及肝损伤指标之间的关系。结果 低值和高值的批内精密度分别为3.01%和2.56%；批间精密度分别为3.74%和2.60%；对接近最低检出限的样本重复检测20次，检出率为100%；此方法学在HBV RNA浓度为1.0×10²～1.0×10⁸拷贝/mL的范围内呈现线性关系(R²=0.996 7)。HBV DNA定量结果低于检测下限组的HBV RNA阳性率为70.73%,HBV DNA未检出组的HBV RNA阳性率为43.68%,2者之间差异具有统计学意义(P=0.004);2组之间的HBV RNA定量检测值差异也具有统计学意义(P=0.000)。HBeAg阳性者的HBV RNA阳性率为89.47%，丙氨酸氨基转移酶(alanine aminotransferase, ALT)异常率57.89%;HBeAg阴性者的HBV RNA阳性率为45.87%,ALT异常率49.54%;2组之间的HBV RNA阳性率差异具有统计学意义(P=0.000)，而ALT异常率之间差异无统计学意义(P=0.502)。HBV RNA阳性的患者ALT异常率为66.7%,HBV RNA检测阴性的患者ALT异常率为37.1%,2者之间差异具有统计学意义(P=0.001)。结论 恒温扩增法检测HBV RNA性能可靠，在HBV DNA低于检测下限的经治CHB患者中仍能有效检测到病毒RNA，可用于该类患者的病情监测。

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 betweenrun 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.