The correlation and consistency of two methods for HBV RNA detection in chronic hepatitis B patients with or without nucleoside/nucleotide analogues treatment

Abstract

Abstract: Objective To compare Race method and Sansure method in HBV RNA quantification in naive and treated patients with chronic hepatitis B (CHB). Methods Clinical datas were collected from patients who were followed up in the outpatient department of Nanfang Hospital. RACE method and Sansure method were used to detect patients' serum HBV RNA levels. The correlation and consistency of the two assays were analyzed. Results A total of 101 patients were enrolled, including 62 patients who received nucleoside/nucleotide analogues (NAs) treatment and 39 patients who did not receive any NAs treatment (i.e., na?ve patients). There were significant differences in HBsAg and HBV DNA levels between the treated and untreated groups. There were statistically differences in serum HBV RNA levels in the two groups. Detectable serum HBV RNA was seen in more patients in RACE group than in Sansure group, as well as higher mean levels of serum HBV RNA (3.16 lg copies/mL vs 2.53 lg copies/mL). There was no significant correlation between HBV DNA levels and HBV RNA levels in the treatment group (RACE R²= 0.4406, Sansure R²= 0.5081); while in the untreated group, there has significant correlation between the two methods (Race R²= 0.8448, Sansure R²= 0.8667). RACE and Sansure were significantly correlated in detectable HBV RNA levels (R²= 0.93498740, P<0.001). In addition, Bland-Altman analysis showed that the 95% of agreement of the two HBV RNA detection methods was relatively narrow (-1.789~0.22 lg copies /mL in the treatment group,and -2.25~0.53 lg copies /mL in the untreated group). If RACE method was used as the baseline, the deviation of Sansure method was -0.78 lg 拷贝/mL in the treatment group and -0.86 lg 拷贝/mL in the untreated group. Linear regression equation: Y (Sansure) =0.918 9X (RACE)-0.3754. Conclusion The correlation between the two methods is significant and can be calculated by linear regression equation; Both the two HBV RNA detection methods can effectively make up for the deficiency of serum HBV DNA detection in guiding the clinical diagnosis and treatment of CHB.

Key words: Hepatitis B virus, HBV RNA, RACE method, Sansure method, Correlation, Consistency

WANG Xue-gang, ZHANG Xin-zhi, YANG Shu, GAN Su-qin, LIU Shi, HAO Kun-yan, ZHOU Bin, YU Yue-cheng. The correlation and consistency of two methods for HBV RNA detection in chronic hepatitis B patients with or without nucleoside/nucleotide analogues treatment[J]. Chinese Hepatolgy, 2020, 25(7): 736-742.

References

[1] 中华医学会感染病学分会,中华医学会肝病学分会. 慢性乙型肝炎防治指南(2019年版). 中华传染病杂志, 2019,37(12): 711-736.
[2] Hou J, Wang G, Wang F, et al. Guideline of Prevention and Treatment for Chronic Hepatitis B (2015 Update). J Clin Transl Hepatol, 2017, 5(4): 297-318.
[3] 高志良. 慢性乙型肝炎功能性治愈(临床治愈)的热点和难点. 现代消化及介入诊疗, 2019, 24(9): 插1, 949-951.
[4] Lai KN, Ho RT, Tam JS, et al. Detection of hepatitis B virus DNA and RNA in kidneys of HBV related glomerulonephritis. Kidney Int, 1996, 50(6): 1965-1977.
[5] Sallie R. Selective detection of hepatitis B virus RNA by PCR. PCR Methods Appl, 1994, 3(6): 376-377.
[6] Gao Y, Li Y, Meng Q, et al. Serum Hepatitis B Virus DNA, RNA, and HBsAg: Which Correlated Better with Intrahepatic Covalently Closed Circular DNA before and after Nucleos(t)ide Analogue Treatment? J Clin Microbiol, 2017, 55(10): 2972-2982.
[7] Wang J, Shen T, Huang X, et al. Serum hepatitis B virus RNA is encapsidated pregenome RNA that may be associated with persistence of viral infection and rebound. J Hepatol, 2016, 65(4): 700-710.
[8] Jansen L, Kootstra NA, van Dort KA, et al. Hepatitis B Virus Pregenomic RNA Is Present in Virions in Plasma and Is Associated With a Response to Pegylated Interferon Alfa-2a and Nucleos(t)ide Analogues. J Infect Dis,2015, 213(2): 224-232.
[9] van Bommel F, Bartens A, Mysickova A, et al. Serum hepatitis B virus RNA levels as an early predictor of hepatitis B envelope antigen seroconversion during treatment with polymerase inhibitors. Hepatology, 2015, 61(1): 66-76.
[10] Tsuge M, Murakami E, Imamura M, et al. Serum HBV RNA and HBeAg are useful markers for the safe discontinuation of nucleotide analogue treatments in chronic hepatitis B patients. J Gastroenterol, 2013, 48(10): 1188-1204.
[11] Huang Y, Chayama K, Tsuge M, et al. Differential effects of interferon and lamivudine on serum HBV RNA inhibition in patients with chronic hepatitis B. Antivir Ther, 2010, 15(2): 177-184.
[12] Liu S, Zhou B, Valdes JD, et al. Serum Hepatitis B Virus RNA: A New Potential Biomarker for Chronic Hepatitis B Virus Infection. Hepatology, 2019, 69(4): 1816-1827.
[13] Bai L, Zhang X, Kozlowski M, et al. Extracellular Hepatitis B Virus RNAs Are Heterogeneous in Length and Circulate as Capsid-Antibody Complexes in Addition to Virions in Chronic Hepatitis B Patients. J Virol, 2018, 92(24).
[14] Zhang W, Hacker H J, Mildenberger M, et al. Detection of HBV RNA in serum of patients. Methods Mol Med, 2004, 95: 29-40.
[15] Huang Q, Zhou B, Cai D, et al. Rapid Turnover of HBV cccDNA Indicated by Monitoring Emergence and Reversion of Signature-Mutation in Treated Chronic Hepatitis B Patients. Hepatology, 2020.
[16] Butler EK, Gersch J, Mcnamara A, et al. Hepatitis B Virus Serum DNA andRNA Levels in Nucleos(t)ide Analog-Treated or Untreated Patients During Chronic and Acute Infection. Hepatology, 2018, 68(6): 2106-2117.
[17] Saldanha J, Gerlich W, Lelie N, et al. An international collaborative study to establish a World Health Organization international standard for hepatitis B virus DNA nucleic acid amplification techniques. Vox Sang, 2001, 80(1): 63-71.
[18] Halgand B, Desterke C, Rivière L, et al. Hepatitis B Virus Pregenomic RNA in Hepatocellular Carcinoma: A Nosological and Prognostic Determinant. Hepatology, 2018, 67(1): 86-96.
[19] Wang J, Yu Y, Li G, et al. Relationship between serum HBV-RNA levels and intrahepatic viral as well as histologic activity markers in entecavir-treated patients. J Hepatol, 2018, 68(1): 16-24.