| Objective:To observe the dynamic change regularity of serum pgRNA during nucleoside analogues(acid)(NAs)antiviral therapy,and the correlation between serum pgRNA and HBsAg,HBV DNA,intrahepatic cccDNA.Besides,to explore the feasibility of serum pgRNA served as the serum maker of cccDNA transcription activity.METHODS:19 CHB/LC patients with complete virological response and maintained response for at lest 3 years were analyzed in this study.Intrahepatic cccDNA was determined by plasmid-safe ATP-dependent Danes(PSAD)digestion in combination with RCA and gap-spanning selective real-time PCR assay after extracted from liver biopsy samples.Human housekeeping genes beta 2-M was used as the reference gene for hepatocyte counting.Quantitative HBsAg levels were determined using chemiluminescence's reagent manufactured by Abbott Company.Serum HBVDNA and pgRNA were quantitied by real-time polymerase chain reaction and real-time reverse transcription polymerase chain reaction assays respectively.Compare the change trend of the three serum makers.Bivariate correlation analysis was applied to analysis the correlation of serum pgRNA and HBs Ag.Compare the ability of serum pgRNA and HBsAg to reflect the cccDNA transcription activity according to the sensitivity ? specificity ?coincidence rate and area under the ROC curve(AUC).RESULTS:(1)During NAs-therapy the levels of all the three serum markers(HBV DNA,pgRNA,HBsAg)were declining.The levels of HBV DNA declined fastest,and pgRNA declined slowly that even maintained virological suppression for 3 years there were still 57.89%(11/19)of patients with positive serum pgRNA(?500 copies/ml),and HBs Ag went slowest that none of the 19 patients met HBsAg negative conversion during this follow-up study.(2)On the base-line,there was a significant correlation(r=0.623,P=0.004)between the levels of serum pgRNA and HBs Ag,besides the overall change trend of the two serum makers was the same during the 3 years of virological suppression,however,when mentioned individual change trend,there were only 47.37%(9/19)of patients had the same serum pgRNA and HBsAg change trend,another 52.63%(10/19)of patients were with different change trend,more over,there were no more significant correlation between serum HBsAg and pgRNA after virologial supression.(3)The coincidence rate of cccDNA and serum pgRNA was 73.68%(14/19),and the coincidence rate of cccDNA and HBsAg was 84.21%(16/19).The sensitivity,specificity,AUC of using serum pgRNA and HBs Ag to speculate if there were residual intrahepatic cccDNA were 68.75%,100%,0.84 and 100%,0,0.64,respectively.(4)The detecting results of cccDNA and serum pgRNA had three models.ModelA : 11 patients(57.89%)had both detectable cccDNA and detectable serum pgRNA.Model B:5 patients(26.32%)had detectable cccDNA but undetectable serum pgRNA.Model C:3 patients(15.79%)had both undetectable cccDNA and undetectable serum pgRNA.In the three models,the levels of cccDNA declined from model A to model C,and only model A had detectable serum pgRNA,but all the three models had high levels HBsAg.CONCLUSION:(1)The ability of serum pgRNA to reflect the level of intrahepatic cccDNA and cccDNA transcriptional activity was better than HBsAg.Besides serum pgRNA was still detectable after HBV DNA was suppressed to undetectable levels,which highlights that it may be a better serum marker in monitoring the effect of NAs therapy.(2)Detecting serum pgRNA is simple and noninvasive,so using pgRNA as the serum marker of monitoring cccDNA transcription activity during NAs therapy has wide prospect of clinical application.(3)Continuous serum pgRNA below the detection limit is expected to become the new index for safe drug withdrawal. |
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