Comparison of fluorescence quantitative PCR and direct PCR sequencing in the detection of HBV polymerase RT region / 中华肝脏病杂志

ABSTRACT

<p><b>OBJECTIVE</b>To explore the feasibility of Direct PCR sequencing in clinic and the significance of Direct PCR sequencing for retrieval treatment plan.</p><p><b>METHODS</b>To address this issue, a cross-sectional study on the drug resistance in the HBV polymerase RT region was performed using Fluorescence quantitative PCR and Direct PCR sequencing in 60 chronic hepatitis B patients, who responded failure to long-term LAM and/or ADV therapy.</p><p><b>RESULT</b>Compared with Fluorescence quantitative PCR, Direct PCR sequencing expressed a higher positive rate (43.2% vs 38.6%; 66.7% vs 54.2%, respectively) and a lower missing rate (9.5% vs 19.0%; 5.9% vs 23.5%, respectively) in the detection of drug resistance in the patients treated with LAM or combination therapy of LAM with ADV. The sensitivity of experiment using Direct PCR sequencing seemed better than Fluorescence quantitative PCR, although the difference was not significant. Further analysis on sensitivity and specificity of detection between high and low viral loads groups indicated that the consensus of two experiments in high viral load group is better than that in low viral load group (Chi-square test = 5.18, probability value = 0.023). In low viral load group, Direct PCR sequencing expressed higher sensitivity and higher specificity than Fluorescence quantitative PCR, although the difference did not approach significant.</p><p><b>CONCLUSION</b>Direct PCR sequencing is better than Fluorescence quantitative PCR in the detection of drug resistance in clinic, not only with higher sensitivity and specificity, but also with comprehensive information of HBV polymerase RT region.</p>