慢性乙型肝炎患者血清HBV恩替卡韦耐药基因突变模式调查*

doi:10.3969/j.issn.1672-5069.2022.03.009

摘要/Abstract

摘要： 目的探讨核苷(酸)类经治的慢性乙型肝炎(CHB)患者血清恩替卡韦耐药基因突变模式的变化。方法 2016年10月～2020年10月我院收治的核苷(酸)类经治的CHB患者107例,采用巢氏PCR法扩增血清HBV RT基因,应用MEGA4软件鉴定病毒基因型。采用脂质体体外转染HepG2细胞,分别给予恩替卡韦、拉米夫定、阿德福韦或替诺福韦作用,采用实时荧光定量PCR法检测培养上清HBV DNA载量。结果在本组107例CHB患者中,恩替卡韦耐药基因rtS202G/M204V/L180M检出率为40.2%,显著高于rtT184A/M204V/L180M(22.2%)、rtT184L/M204V/L180M(23.4%)、rtS202G/M204i/L180M(3.7%)、rtM250V/M204V/L180M(5.6%)、rtM250L/M204I/L180M(5.6%)和rtT184I/S202G/M204V/L180M(5.6%)等基因型(P<0.05);恩替卡韦耐药患者曾经拉米夫定治疗率为38.3%,显著高于应用过阿德福韦治疗者的22.4%(P<0.05); 36例rtT184^sub耐药基因感染者血清HBV DNA为(3.0±0.5)lgIU/ml,10例rtT184/S202^sub感染者血清HBV DNA为(4.1±0.8) lgIU/ml,显著高于47例rtS202^sub感染者【(1.9±0.2)lgIU/ml,P<0.05】或14例rtM250^sub感染者【(2.1±0.2)lgIU/ml,P<0.05】;体外在恩替卡韦作用的携带rtS202G/M204V/L180M突变株细胞培养上清HBV DNA为(3.1±0.1)lg IU/ml,显著高于rtT184A/M204V/L180M突变株【(2.1±0.1 lg IU/ml,P<0.05】,而在拉米夫定或阿德福韦酯作用细胞,培养上清HBV DNA水平无显著性差异(P>0.05),在替诺福韦作用细胞培养上清,HBV DNA水平均很低。结论核苷(酸)经治CHB患者恩替卡韦耐药突变基因型主要为rtS202G/M204V/L180M,变异株对拉米夫定、阿德福韦酯和恩替卡韦抵抗,但对替诺福韦仍敏感。

关键词: 慢性乙型肝炎, 核苷(酸)类, 经治, 耐药基因突变, 替诺福韦

Abstract: Objective The aim of this study was to investigate entecavir resistance gene mutation in nucleosi(t)de-treated patients with chronic hepatitis B (CHB). Methods 107 nucleosi(t)de-treated patients with CHB and entecavir-resistance mutation were enrolled in our hospital between October 2016 and October 2020. Serum gene amplification of HBV RT was conducted by nested PCR, and the genotypes were identified by MEGA4 software. The HepG2 cells were transfected by liposome in vitro with different HBV mutants, the cells were intervened by lamivudine, adefovir, entecavir and tenofovir, and the supernatant HBV DNA loads were detected by PCR. Results Out of the 107 nucleosi(t)de-treated patients with CHB in our series, the percentages of entecavir resistance gene rtS202G/M204V/L180M was 40.2%, significantly higher than in patients with rtT184A/M204V/L180M(22.2%), rtT184L/M204V/L180M(23.4%), rtS202G/M204i/L180M(3.7%), rtM250V/M204V/L180M(5.6%), rtM250L/M204I/L180M(5.6%) and rtT184I/S202G/M204V/L180M(5.6%) infections (P<0.05); 38.3% patients had received lamivudine, significantly higher than 22.4% receiving adefovir therapy in our series(P<0.05); serum HBV DNA loads in 36 patients with rtT184^sub mutant infection was (3.0±0.5)lgIU/ml, and in 10 with rtT184/S202^sub infection was (4.1±0.8) lgIU/ml, both significantly higher than [(1.9±0.2)lgIU/ml, P<0.05] in 47 patients with rtS202^sub infection or [(2.1±0.2)lgIU/ml, P<0.05] in 14 patients with rtM250^sub infection; the supernatant HBV DNA load in entecavir-intervened cells carrying rtS202G/M204V/L180M mutant was (3.1±0.1)lg IU/ml, significantly higher than [(2.1±0.1 lg IU/ml, P<0.05] in cells carrying rtT184A/M204V/L180M mutants, no significant differences in lamivudine- or adefovir-treated cells(P>0.05), and they both decreased to undetectable in tenofovir-intervened cells. Conclusion The superior entecavir-resistance mutation in nucleosi(t)de-treated patients with CHB is rtS202G/M204V/L180M infection, and they are resistant to lamivudine or adefovir, but fortunately, they still sensitive to tenofovir intervention.

Key words: Hepatitis B, Nucleosi(t)de-treatment, Drug resistance gene, Mutant infection, Tenofovir

王广丽, 徐欢, 董丹丹, 黄荷. 慢性乙型肝炎患者血清HBV恩替卡韦耐药基因突变模式调查^*[J]. 实用肝脏病杂志, 2022, 25(3): 339-342.

Wang Guangli, Xu Huan, Dong Dandan, et al. Entecavir resistance gene mutation in nucleosi(t)de-treated patients with chronic hepatitis B[J]. Journal of Practical Hepatology, 2022, 25(3): 339-342.

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慢性乙型肝炎患者血清HBV恩替卡韦耐药基因突变模式调查^*

Entecavir resistance gene mutation in nucleosi(t)de-treated patients with chronic hepatitis B

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